Monoclonal antibody-cytokine fusion protein dimer and application thereof

ABSTRACT

Provided are a monoclonal antibody-cytokine fusion protein dimer and an application thereof. The monoclonal antibody comprises two heavy chains, the fusion protein dimer comprises a first heavy chain polypeptide chain and a second heavy chain polypeptide chain, wherein one heavy chain of the monoclonal antibody is connected to a cytokine to form the first heavy chain polypeptide chain, and the other heavy chain of the monoclonal antibody is optionally connected to another cytokine to form the second heavy chain polypeptide chain. The monoclonal antibody-cytokine fusion protein dimer provided not only fully retains the biological activity of the antibody, but also significantly improves the biological activity of the cytokine(s). More importantly, by using the target specificity of the antibody molecule, the immune response of immune cells is enhanced and the toxicity of the cytokine(s) is reduced, and thus, on the premise of enhancing the efficacy of the drug, the safety of the medication is ensured.

TECHNICAL FIELD

The present invention relates to a fusion protein, especially a protein dimer based on monoclonal antibody and cytokine fusion and an application thereof.

BACKGROUND

An immune system is a set of defense system in mammal bodies and can resist invasion of various pathogens and eliminate tumors. Antibody molecules play an important role in the immune system, for example, recognizing a tumor antigen and activating an immune response. Meanwhile, cytokines serve as immunomodulatory factors to be capable of regulating and controlling innate immunity and adaptive immunity at the same time, and also have significance which cannot be ignored in cancer immunotherapy. This kind of regulatory factors almost participate in various links of the immune system, and perform a powerful function in aspects of regulating and balancing the immune system. Interferon and IL-2 are first selected as cytokines to be applied to immunotherapy. IFN-α is used to treat various cancers, such as melanoma, renal cancer, follicular lymphoma, and chronic myelogenous leukemia, while IL-2 was approved by FDA in 1998 to be used to treat advanced metastatic melanoma, metastatic renal cancer and the like. Other cytokines, such as IL-7, IL-10, IL-12, IL-15 and IL-21, are also in a clinical testing stage. Therefore, the cytokines have a certain potential to be used in immunotherapy.

However, the immunotherapy based on the cytokines is severely hindered in clinical use, and is mainly enslaved to severe side effects and underperformed pharmacokinetic properties. Therefore, in order to improve the treatment effect of the cytokines, the cytokines frequently need to be transformed. At present, a common method is to adopt a method of antibody fusion, and the cytokines are targetedly delivered to specific parts through the antibody, thereby effectively reducing cytotoxicity, improving the pharmacokinetic properties, and then enhancing an immunoregulation function. Even so, after some cytokines are fused onto the antibody, the activity of the antibody will be reduced, thereby influencing an overall effect. In addition, some cytokines can only exert the respective optimal effect by combined use, but the prior art cannot meet this requirement.

In immunoregulation, activating and inhibiting an immune response are mainly regulated by two independent signaling pathways. A first signal is antigen-mediated. When a T cell receptor specifically recognizes and binds to an antigen peptide presented by MHC on surfaces of antigen-presenting cells (APC), the first signal is generated. A second signal is provided by the interaction between antigen-presenting cells and costimulatory molecules expressed on the surface of T cells. When the first and second signals are activated in sequence, the tumors can be killed by T cells. If the second signal is in lack, T cells will enter an unresponsive state or immune tolerance, and even cause programmed cell death.

As mentioned above, the second signaling pathway is very important for activating immune cells. Specifically, co-stimulatory and co-inhibitory receptors participate in the second signaling pathway, induce an immune response and regulation of antigen-receptor presentation, balance positive and negative signals while maintaining immune tolerance of autoantigens, and maximize the immune response to invaders. CD28 is a member of a CD28 family, is a major T cell costimulatory receptor, and is subjected to constitutive expression on initial CD4+ and CD8+ T cells. Cytotoxic T lymphocyte-associated antigen 4 (CTLA-4) molecules are also a member of the CD28 receptor family, but are coinhibitory receptors, and the molecules are subjected to constitutive expression on regulatory T cells (Treg) or are generated after the T cells are activated by CD28. Either CD28 or CTLA-4 can be combined with a B7.1 (CD80) or B7.2 (CD86) ligand so as to deliver activating or inhibiting signals in the T cells respectively.

Ligands of the CD28 receptor include CD80, CD86, a programmed death ligand 1 (PD-L1) and a programmed death ligand 2 (PD-L2). Among these is PD-L1, a transmembrane protein that binds to the inhibitory checkpoint molecule of PD-1 and suppresses the adaptive immune response by delivering an inhibitory “don't find me” signal. A PD-1/PD-L1 signaling pathway plays a critical role in development of immune tolerance, and prevents an overreaction of the immune system so as to avoid an autoimmune disease. However, it is usually deregulated in a cancer progress, and tumor cells are allowed to disguise as healthy tissues to circumvent a protection mechanism. The tumor cells with overexpression of PD-L1 can escape T cell mediated death by activating the PD-1/PD-L1 inhibitory signaling pathway so as to inhibit an anti-tumor adaptive immune reaction. PD-1 overexpression in human tumor-associated macrophages (TAMS) is also proved to be capable of inhibiting phagocytosis and tumor immunity. At present, an anti-PD-1 or anti-PD-L1 monoclonal antibody for blocking PD-1/PD-L1 interaction has remarkable effect in cancer treatment. Although Food and Drug Administration approves the anti-PD-1 or anti-PD-L1 monoclonal antibody such as keytruda, opdivo and tecentriq to be used to treat advanced cancers, these anti-tumor drugs only have reaction on part of patients, and it implies that single blocking of the inhibitory signaling pathway is insufficient to activate an immune response, and there are still other mechanisms to inhibit the immune system.

A lymphocyte activation gene 3 (LAG-3) is a transmembrane protein, and is expressed in the activated T cells, natural killer cells, B cells and plasmacytoid dendritic cells. Like PD-1, LAG-3 is one of immune checkpoint receptors combined with MHC II and negatively regulating T cell receptor signal transduction on APC. Because LAG-3 is also expressed on the Treg cells, LAG-3 blocking can inhibit the activity of the Treg cells and enhance its anti-tumor immune function. Preclinical experiments show that LAG-3 signaling pathway blocking has the anti-tumor effect.

TIGIT, known as T cell immunoglobulin and ITIM domain protein, is an inhibitory receptor containing Ig and ITIM domains shared by T cells and NK cells. TIGIT is highly expressed in T cells and natural killer (NK) cells. TIGIT, CD96, CD226 and related ligands together form an immunomodulatory signaling pathway. Similar to the CD28/CTLA-4 signaling pathway, the CD226/TIGIT/CD96 signaling pathway also contains costimulatory receptors and coinhibitory receptors sharing some or all of the ligands, in which CD226 is a costimulatory receptor and delivers stimulus signals upon combining with ligands, and TIGIT and CD96 are coinhibitory receptors and deliver inhibitory signals upon combining with related ligands. TIGIT has two ligands, i.e. CD155 and CD122, which are also ligands of CD226. These two ligands are expressed in APC cells, T cells and tumor cells. The ligands of CD96 include CD155 and CD111. The affinity of TIGIT with the ligand CD155 is significantly higher than that of TIGIT with the ligand CD122, and also significantly higher than the affinity of CD226 or CD96 with the ligand CD155. Similar to PD-1 and CTLA-4 receptors, TIGIT is also an important inhibitory immune receptor. Inhibition of TIGIT can promote the proliferation and function of T cells; blocking of TIGIT can also enhance the anti-tumor immune response mediated by NK cells, thereby inhibiting tumor growth. Therefore, monoclonal antibodies targeting the inhibitory receptor TIGIT can significantly enhance the effect of tumor immunotherapy.

Currently, with the successful launch of PD-1/PD-L1 antibodies, immunotherapy based on immune checkpoint blockade has been rapidly developed and promoted. However, these antibodies are only effective to a part of patients, and will generate drug resistance. Therefore, how to better combine the cytokines and the antibodies to solve these problems is always a research focus.

SUMMARY

In order to solve the above problems, in one aspect, the present invention provides a monoclonal antibody-cytokine fusion protein dimer, the monoclonal antibody includes two heavy chains, the fusion protein dimer includes a first heavy chain polypeptide chain and a second heavy chain polypeptide chain, where one heavy chain of the monoclonal antibody is connected with a cytokine to form the first heavy chain polypeptide chain, and the other heavy chain of the monoclonal antibody is optionally connected with another cytokine to form the second heavy chain polypeptide chain.

In some embodiments, the monoclonal antibody is an anti-immune checkpoint molecular antibody. Preferably, the monoclonal antibody is selected from an anti-LAG-3 antibody, an anti-PD-L1 antibody, an anti-TIGIT antibody or an anti-PD-1 antibody. In some embodiments, the monoclonal antibody is the anti-LAG-3 antibody. In some other embodiments, the monoclonal antibody is the anti-PD-L1 antibody. In some embodiments, the monoclonal antibody is the anti-TIGIT antibody. In some other embodiments, the monoclonal antibody is the anti-PD-1 antibody.

In some embodiments, the two cytokines are selected from IL-2, IL-12, GM-CSF, an IL-2 mutant and a combination thereof. Preferably, the two cytokines are selected from IL-2, IL-12 or the IL-2 mutant or a combination thereof.

In some embodiments, two subunits P35 and P40 of IL-12 are connected through a linker sequence to form an IL-12 single strand protein so as to exist in the fusion protein dimer. In some specific embodiments, the two subunits P35 and P40 of IL-12 are connected through the linker sequence to form the IL-12 single strand protein connected with one heavy chain of the monoclonal antibody to form the first or second heavy chain polypeptide chain.

In some embodiments, the monoclonal antibody and the cytokines are connected through the linker sequence or directly fused. In some embodiments, the linker sequence is selected from (S(G4S)₁₋₃), (G4S)₁₋₃, KRVAPELLGGPS, ASTKG, NSPPAA, EPKSSDKTHTSPPSP, ERKSSVESPPS and ESKYGPPSPPSP. Preferably, the linker sequence is (S(G4S)₁₋₃). More preferably, the linker sequence is (S(G4S)₃) (SEQ ID NO:63).

In some embodiments, the monoclonal antibody is connected with the cytokines through Fc regions of the heavy chains. In some embodiments, the Fc regions of the two heavy chains of the monoclonal antibody are different and have an asymmetric complementary structure therebetween. The asymmetric complementary structure may be, for example, formed through a knobs-into-holes (KiH) technology. Preferably, the Fc regions of the two heavy chains of the monoclonal antibody have mutation combinations selected from a mutation combination T366W/S354C and a mutation combination T366S/L368A/Y407V/Y349C. In some specific embodiments, the Fc region of any heavy chain of the monoclonal antibody has the mutation combination T366W/S354C, and the Fc region of the other complementary heavy chain has the mutation combination T366S/L368A/Y407V/Y349C. In some specific embodiments, the Fc regions of the heavy chains of the monoclonal antibody are selected from a human IgG1 mutant and a human IgG4 mutant.

In some embodiments, one heavy chain of the monoclonal antibody is connected with the cytokine IL-12 to form the first heavy chain polypeptide chain, and the other heavy chain of the monoclonal antibody is connected with the cytokine IL-2 or IL-2 mutant to form the second heavy chain polypeptide chain; or one heavy chain of the monoclonal antibody is connected with the cytokine IL-2 or IL-2 mutant to form the first heavy chain polypeptide chain, and the other heavy chain of the monoclonal antibody is connected with the cytokine IL-12 to form the second heavy chain polypeptide chain. In some embodiments, one heavy chain of the monoclonal antibody is connected with the cytokine IL-12, IL-2 or IL-2 mutant to form the first heavy chain polypeptide chain, and the other heavy chain of the monoclonal antibody is not connected with the cytokine to serve as the second heavy chain polypeptide chain. In some embodiments, one heavy chain of the monoclonal antibody is connected with the cytokine IL-12 to form the first heavy chain polypeptide chain, and the other heavy chain of the monoclonal antibody is not connected with the cytokine to serve as the second heavy chain polypeptide chain. In some embodiments, one heavy chain of the monoclonal antibody is connected with the cytokine IL-2 or IL-2 mutant to form the first heavy chain polypeptide chain, and the other heavy chain of the monoclonal antibody is not connected with the cytokine to serve as the second heavy chain polypeptide chain.

In some embodiments, the cytokines are connected to a C-terminus of the heavy chain Fc regions of the monoclonal antibody through the linker sequence. In some other embodiments, the cytokines are directly connected to the C-terminus of the heavy chain Fc regions of the monoclonal antibody.

In some embodiments, a heavy chain of the anti-LAG-3 monoclonal antibody has a sequence with at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identity with an amino acid sequence shown in SEQ ID NOs:8, 12 or 14, and a light chain of the anti-LAG-3 monoclonal antibody has a sequence with at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identity with an amino acid sequence shown in SEQ ID NO:10. In some specific embodiments, the heavy chain of the anti-LAG-3 monoclonal antibody has the amino acid sequence as shown in SEQ ID NOs:8, 12 or 14, and the light chain of the anti-LAG-3 monoclonal antibody has the amino acid sequence as shown in SEQ ID NO:10.

In some embodiments, a heavy chain of the anti-PD-L1 monoclonal antibody has a sequence with at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identity with an amino acid sequence shown in SEQ ID NOs:22, 26 or 28, and a light chain of the anti-PD-L1 monoclonal antibody has a sequence with at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identity with an amino acid sequence shown in SEQ ID NO:24. In some embodiments, the heavy chain of the anti-PD-L1 monoclonal antibody has the amino acid sequence as shown in SEQ ID NOs:22, 26 or 28, and the light chain of the anti-PD-L1 monoclonal antibody has the amino acid sequence as shown in SEQ ID NO:24.

In some embodiments, a heavy chain of the anti-TIGIT monoclonal antibody has a sequence with at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identity with an amino acid sequence shown in SEQ ID NOs:36, 40 or 42, and a light chain of the anti-TIGIT monoclonal antibody has a sequence with at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identity with an amino acid sequence shown in SEQ ID NO:38. In some embodiments, the heavy chain of the anti-TIGIT monoclonal antibody has the amino acid sequence as shown in SEQ ID NOs:36, 40 or 42, and the light chain of the anti-TIGIT monoclonal antibody has the amino acid sequence as shown in SEQ ID NO:38.

In some embodiments, a heavy chain of the anti-PD-1 monoclonal antibody has a sequence with at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identity with an amino acid sequence shown in SEQ ID NOs:50, 54 or 56, and a light chain of the anti-PD-1 monoclonal antibody has a sequence with at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identity with an amino acid sequence shown in SEQ ID NO:52. In some embodiments, the heavy chain of the anti-PD-1 monoclonal antibody has the amino acid sequence as shown in SEQ ID NOs:50, 54 or 56, and the light chain of the anti-PD-1 monoclonal antibody has the amino acid sequence as shown in SEQ ID NO:52.

In some embodiments, the IL-12 single strand protein has an amino acid sequence as shown in SEQ ID NO:4.

In some embodiments, IL-2 has an amino acid sequence as shown in SEQ ID NO:8.

The fusion protein dimer provided by the present invention includes two light chains of the monoclonal antibody.

In some embodiments, the first and second heavy chain polypeptide chains of the fusion protein are selected from sequences with at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identity with an amino acid sequence shown in SEQ ID NOs:12, 14, 16, 18, 20, 26, 28, 30, 32, 34, 40, 42, 44, 46, 48, 54, 56, 58, 60 or 62. In some specific embodiments, the first and second heavy chain polypeptide chains of the fusion protein are selected from an amino acid sequence as shown in SEQ ID NOs:12, 14, 16, 18, 20, 26, 28, 30, 32, 34, 40, 42, 44, 46, 48, 54, 56, 58, 60 or 62.

In some embodiments, the monoclonal antibody is the anti-LAG-3 antibody, and the first and second heavy chain polypeptide chains are selected from sequences with at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identity with an amino acid sequence shown in SEQ ID NOs:12, 14, 16, 18 or 20. In some specific embodiments, the monoclonal antibody is the anti-LAG-3 antibody, and the first and second heavy chain polypeptide chains are selected from the amino acid sequence shown in SEQ ID NOs:12, 14, 16, 18 or 20. In some specific embodiments, the monoclonal antibody is the anti-LAG-3 antibody, where the first heavy chain polypeptide chain has the amino acid sequence as shown in SEQ ID NO:18, and the second heavy chain polypeptide chain has the amino acid sequence as shown in SEQ ID NO:16; the first heavy chain polypeptide chain has the amino acid sequence as shown in SEQ ID NO:16, and the second heavy chain polypeptide chain has the amino acid sequence as shown in SEQ ID NO:14; or the first heavy chain polypeptide chain has the amino acid sequence as shown in SEQ ID NO:20, and the second heavy chain polypeptide chain has the amino acid sequence as shown in SEQ ID NO:14.

In some embodiments, the present invention provides an anti-LAG-3 monoclonal antibody-cytokine fusion protein dimer, including a first heavy chain polypeptide chain, a second heavy chain polypeptide chain and two light chains of an anti-LAG-3 antibody, where the first heavy chain polypeptide chain has the amino acid sequence as shown in SEQ ID NO:18, the second heavy chain polypeptide chain has the amino acid sequence as shown in SEQ ID NO:16, and the light chains have the amino acid sequence as shown in SEQ ID NO:8. In some other embodiments, the present invention provides an anti-LAG-3 monoclonal antibody-cytokine fusion protein dimer, including a first heavy chain polypeptide chain, a second heavy chain polypeptide chain and two light chains of an anti-LAG-3 antibody, where the first heavy chain polypeptide chain has the amino acid sequence as shown in SEQ ID NO:16, the second heavy chain polypeptide chain has the amino acid sequence as shown in SEQ ID NO:14, and the light chains have the amino acid sequence as shown in SEQ ID NO:8. In another embodiment, the present invention provides an anti-LAG-3 monoclonal antibody-cytokine fusion protein dimer, including a first heavy chain polypeptide chain, a second heavy chain polypeptide chain and two light chains of an anti-LAG-3 antibody, where the first heavy chain polypeptide chain has the amino acid sequence as shown in SEQ ID NO:20, the second heavy chain polypeptide chain has the amino acid sequence as shown in SEQ ID NO:14, and the light chains have the amino acid sequence as shown in SEQ ID NO:8.

In some embodiments, the monoclonal antibody is the anti-PD-L1 antibody, and the first and second heavy chain polypeptide chains are selected from sequences with at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identity with an amino acid sequence shown in SEQ ID NOs:26, 28, 30, 32 or 34. In some specific embodiments, the monoclonal antibody is the anti-PD-L1 antibody, and the first and second heavy chain polypeptide chains are selected from the amino acid sequence shown in SEQ ID NOs:26, 28, 30, 32 or 34. In some specific embodiments, the monoclonal antibody is the anti-PD-L1 antibody, where the first heavy chain polypeptide chain has the amino acid sequence as shown in SEQ ID NO:32, and the second heavy chain polypeptide chain has the amino acid sequence as shown in SEQ ID NO:30; the first heavy chain polypeptide chain has the amino acid sequence as shown in SEQ ID NO:30, and the second heavy chain polypeptide chain has the amino acid sequence as shown in SEQ ID NO:28; or the monoclonal antibody is the anti-PD-L1 antibody, where the first heavy chain polypeptide chain has the amino acid sequence as shown in SEQ ID NO:34, and the second heavy chain polypeptide chain has the amino acid sequence as shown in SEQ ID NO:28.

In some embodiments, the present invention provides an anti-PD-L1 monoclonal antibody-cytokine fusion protein dimer, including a first heavy chain polypeptide chain, a second heavy chain polypeptide chain and two light chains of an anti-PD-L1 antibody, where the first heavy chain polypeptide chain has the amino acid sequence as shown in SEQ ID NO:32, the second heavy chain polypeptide chain has the amino acid sequence as shown in SEQ ID NO:30, and the light chains have the amino acid sequence as shown in SEQ ID NO:24. In some other embodiments, the present invention provides an anti-PD-L1 monoclonal antibody-cytokine fusion protein dimer, including a first heavy chain polypeptide chain, a second heavy chain polypeptide chain and two light chains of an anti-PD-L1 antibody, where the first heavy chain polypeptide chain has the amino acid sequence as shown in SEQ ID NO:30, the second heavy chain polypeptide chain has the amino acid sequence as shown in SEQ ID NO:28, and the light chains have the amino acid sequence as shown in SEQ ID NO:24. In some more embodiments, the present invention provides an anti-PD-L1 monoclonal antibody-cytokine fusion protein dimer, including a first heavy chain polypeptide chain, a second heavy chain polypeptide chain and two light chains of an anti-PD-L1 antibody, where the first heavy chain polypeptide chain has the amino acid sequence as shown in SEQ ID NO:34, the second heavy chain polypeptide chain has the amino acid sequence as shown in SEQ ID NO:28, and the light chains have the amino acid sequence as shown in SEQ ID NO:24.

In some embodiments, the monoclonal antibody is the anti-TIGIT antibody, and the first and second heavy chain polypeptide chains are selected from sequences with at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identity with an amino acid sequence shown in SEQ ID NOs:40, 42, 44, 46 or 48. In some specific embodiments, the monoclonal antibody is the anti-TIGIT antibody, and the first and second heavy chain polypeptide chains are selected from the amino acid sequence shown in SEQ ID NOs:40, 42, 44, 46 or 48. In some specific embodiments, the monoclonal antibody is the anti-TIGIT antibody, where the first heavy chain polypeptide chain has the amino acid sequence as shown in SEQ ID NO:46, and the second heavy chain polypeptide chain has the amino acid sequence as shown in SEQ ID NO:44; the first heavy chain polypeptide chain has the amino acid sequence as shown in SEQ ID NO:44, and the second heavy chain polypeptide chain has the amino acid sequence as shown in SEQ ID NO:42; or the first heavy chain polypeptide chain has the amino acid sequence as shown in SEQ ID NO:48, and the second heavy chain polypeptide chain has the amino acid sequence as shown in SEQ ID NO:42.

In some embodiments, the present invention provides an anti-TIGIT monoclonal antibody-cytokine fusion protein dimer, including a first heavy chain polypeptide chain, a second heavy chain polypeptide chain and two light chains of an anti-TIGIT antibody, where the first heavy chain polypeptide chain has the amino acid sequence as shown in SEQ ID NO:46, the second heavy chain polypeptide chain has the amino acid sequence as shown in SEQ ID NO:44, and the light chains have the amino acid sequence as shown in SEQ ID NO:38. In some other embodiments, the present invention provides an anti-TIGIT monoclonal antibody-cytokine fusion protein dimer, including a first heavy chain polypeptide chain, a second heavy chain polypeptide chain and two light chains of an anti-TIGIT antibody, where the first heavy chain polypeptide chain has the amino acid sequence as shown in SEQ ID NO:44, the second heavy chain polypeptide chain has the amino acid sequence as shown in SEQ ID NO:42, and the light chains have the amino acid sequence as shown in SEQ ID NO:38. In some more embodiments, the present invention provides an anti-TIGIT monoclonal antibody-cytokine fusion protein dimer, including a first heavy chain polypeptide chain, a second heavy chain polypeptide chain and two light chains of an anti-TIGIT antibody, where the first heavy chain polypeptide chain has the amino acid sequence as shown in SEQ ID NO:48, the second heavy chain polypeptide chain has the amino acid sequence as shown in SEQ ID NO:42, and the light chains have the amino acid sequence as shown in SEQ ID NO:38.

In some embodiments, the monoclonal antibody is the anti-PD-1 antibody, and the first and second heavy chain polypeptide chains are selected from sequences with at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identity with an amino acid sequence shown in SEQ ID NOs:54, 56, 58, 60 or 62. In some specific embodiments, the monoclonal antibody is the anti-PD-1 antibody, and the first and second heavy chain polypeptide chains are selected from the amino acid sequence shown in SEQ ID NOs:54, 56, 58, 60 or 62. In some specific embodiments, the monoclonal antibody is the anti-PD-1 antibody, where the first heavy chain polypeptide chain has the amino acid sequence as shown in SEQ ID NO:60, and the second heavy chain polypeptide chain has the amino acid sequence as shown in SEQ ID NO:58; the first heavy chain polypeptide chain has the amino acid sequence as shown in SEQ ID NO:58, and the second heavy chain polypeptide chain has the amino acid sequence as shown in SEQ ID NO:56; or the first heavy chain polypeptide chain has the amino acid sequence as shown in SEQ ID NO:62, and the second heavy chain polypeptide chain has the amino acid sequence as shown in SEQ ID NO:56.

In some embodiments, the present invention provides an anti-PD-1 monoclonal antibody-cytokine fusion protein dimer, including a first heavy chain polypeptide chain, a second heavy chain polypeptide chain and two light chains of an anti-PD-1 antibody, where the first heavy chain polypeptide chain has the amino acid sequence as shown in SEQ ID NO:60, the second heavy chain polypeptide chain has the amino acid sequence as shown in SEQ ID NO:58, and the light chains have the amino acid sequence as shown in SEQ ID NO:52. In some other embodiments, the present invention provides an anti-PD-1 monoclonal antibody-cytokine fusion protein dimer, including a first heavy chain polypeptide chain, a second heavy chain polypeptide chain and two light chains of an anti-PD-1 antibody, where the first heavy chain polypeptide chain has the amino acid sequence as shown in SEQ ID NO:58, the second heavy chain polypeptide chain has the amino acid sequence as shown in SEQ ID NO:56, and the light chains have the amino acid sequence as shown in SEQ ID NO:52. In some more embodiments, the present invention provides an anti-PD-1 monoclonal antibody-cytokine fusion protein dimer, including a first heavy chain polypeptide chain, a second heavy chain polypeptide chain and two light chains of an anti-PD-1 antibody, where the first heavy chain polypeptide chain has the amino acid sequence as shown in SEQ ID NO:62, the second heavy chain polypeptide chain has the amino acid sequence as shown in SEQ ID NO:56, and the light chains have the amino acid sequence as shown in SEQ ID NO:52.

In another aspect, the present invention provides an isolated polynucleotide, encoding a first or second heavy chain polypeptide chain of the fusion protein dimer thereof as described herein.

In some specific embodiments, the polynucleotide has a nucleotide sequence as shown in SEQ ID NO:11, 13, 15, 17, 19, 25, 27, 29, 31, 33, 39, 41, 43, 45, 47, 53, 55, 57, 59 or 61.

In another aspect, the present invention provides an expression vector containing the polynucleotide.

In another aspect, the present invention provides a host cell containing the expression vector.

In another aspect, the present invention provides an application of the fusion protein dimer, polynucleotide, the expression vector or the host cell in the preparation of an anti-tumor drug. In some embodiments, the tumor is melanoma, stomach cancer, renal cancer, Hodgkin lymphoma, head and neck cancer, bladder cancer or non-small cell lung cancer. Preferably, the tumor is melanoma, stomach cancer or renal cancer.

In another aspect, the present invention provides an anti-tumor pharmaceutical composition, where the pharmaceutical composition contains the fusion protein dimer and a pharmaceutically acceptable carrier.

In another aspect, the present invention provides a tumor treating method, comprising administering to the subject a therapeutically effective amount of the fusion protein dimer or the pharmaceutical composition containing the fusion protein dimer.

In some embodiments, the anti-tumor pharmaceutical composition provided by the present invention may be administered to the subject by at least one route selected from the following: parenteral, subcutaneous, intramuscular, intravenous, intra-articular, endobronchial, intra-abdominal, intracapsular, endochondral, endoluminal, intracelial, intracerebellar, intracerebroventricular, intracolonic, intracervical, entogastric, intrahepatic, intramyocardial, endosseous, endopelvic, intrapericardial, intraperitoneal, intrapleural, intraprostatic, intrapulmonary, intrarectal, intrarenal, intraretinal, intraspinal, intrasynovial, intrapleural, intratympanic, intrauterine, intravesical, intravitreous, rapid injection, subconjunctival, transvaginal, transrectal, transbuccal, sublingual, intranasal, intratumoral and transcutaneous. Preferably, the anti-tumor pharmaceutical composition is administered to the subject in an intratumoral or intravenous mode.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 Structure of monoclonal antibody-cytokine fusion protein dimer

FIG. 2 IL-2 activity detection of an anti-LAG-3 antibody-cytokine fusion protein

FIG. 3 IL-2 activity detection of an anti-PD-L1 antibody-cytokine fusion protein

FIG. 4 IL-2 activity detection of an anti-TIGIT antibody-cytokine fusion protein

FIG. 5 IL-2 activity detection of an anti-PD-1 antibody-cytokine fusion protein

FIG. 6 IL-12 activity detection of an anti-LAG-3 antibody-cytokine fusion protein

FIG. 7 IL-12 activity detection of an anti-PD-L1 antibody-cytokine fusion protein

FIG. 8 IL-12 activity detection of an anti-TIGIT antibody-cytokine fusion protein

FIG. 9 IL-12 activity detection of an anti-PD-1 antibody-cytokine fusion protein

FIG. 10 Activity detection of LAG-3 blocking of an anti-LAG-3 antibody-cytokine fusion protein

FIG. 11 Activity detection of PD-L1 blocking of an anti-PD-L1 antibody-cytokine fusion protein

FIG. 12 Activity detection of TIGIT blocking of an anti-TIGIT antibody-cytokine fusion protein

FIG. 13 Activity detection of PD-1 blocking of an anti-PD-1 antibody-cytokine fusion protein

DETAILED DESCRIPTION

Unless otherwise defined, technical or scientific terms used in the present invention shall have the meanings generally understood by a person skilled in the art to which the present invention pertains.

The present invention provides a fusion protein with anti-tumor activity by utilizing an antibody Fc heterodimer technology, and the fusion protein includes a monoclonal antibody and one or more cytokines. In the present invention, “monoclonal antibody-cytokine fusion protein”, “antibody/cytokine fusion protein” and “McAb-cytokine fusion protein” all refer to the monoclonal antibody-cytokine fusion protein based on the antibody Fc heterodimer technology, including that the one or more cytokines are connected through an antibody heavy chain Fc region to form the fusion protein. Optionally, the monoclonal antibody and the cytokines are connected to a C-terminus of the antibody heavy chain Fc region through a linker sequence.

The term “antibody” refers to a large “Y” shaped protein secreted by a plasmacyte (effector B cells) and used by an immune system to identify and neutralize foreign substances such as bacteria and viruses. In the past more than 10 years, more and more monoclonal antibodies are widely applied to tumor treatment. The antibody usually is a tetramer formed by 2 same heavy chains and 2 same light chains which are mutually connected through disulfide bonds. “anti-immune checkpoint molecular antibody” involved in the present invention refers to an antibody specifically binding to immune checkpoint molecules. The immune checkpoint molecules include but not limited to LAG-3, PD-L1, TIGIT, PD-1 or CTLA-4, and the antibody refers to the monoclonal antibody with the two heavy chains and the two light chains.

The term “antibody Fc region” or “antibody Fc” refers to a shank region of a “Y” shape, namely, a fragment crystallizable (Fc), including second and third constant domains (CH2 and CH3 domains) of the heavy chains. The antibody Fc region may be obtained through hydrolyzing antibody molecules by a proteolytic enzyme (such as papain).

A dimer protein involved in the present invention means that in a protein forming process, if two subunits/monomers are the same, it is called a homodimer (homo-), and if the dimer protein is combined by subunits/monomers which are not exactly the same, it is called a heterodimer (hetero-). The term “antibody Fc heterodimer” means that the protein is combined by the two different subunits/monomers, and each subunit/monomer contains an antibody Fc fragment. The key is that the two antibody Fc fragments have different amino acid site mutations, so that complementary protein spatial structures can be formed, and the two different subunits/monomers can be correctly combined.

The term “knobs-into-holes (KiH) technology” refers to a technology for promoting assembly between two kinds of heterologous antibody heavy chains. For example, a site 366 small-size threonine (T) at a heavy chain CH3 region of one antibody may be mutated into a large-size tyrosine (Y) so as to form a protruding “knobs” type structure (T366Y); meanwhile, a site 407 large tyrosine (Y) residue at a heavy chain CH3 region of another antibody is mutated into a small threonine (T) so as to form a recessed “holes” type structure (Y407T). The two kinds of different antibody heavy chains can be correctly assembled by utilizing a steric-hindrance effect of the “knobs-into-holes” structure (namely, an asymmetric complementary structure). According to the present invention, the better assembly effect is achieved by mutation and combination of a plurality of sites in the two antibody Fc regions.

The term “cytokine (CK)” is a low-molecular-weight soluble protein generated by inducing various cells through an immunogen, a mitogen or other stimulants. In an organism, the cytokine is combined with its specific cell surface receptor to deliver intracellular signals so as to change a cell function, and has the various effects of regulating innate immunity and adaptive immunity, hemocytogenesis, cell growth, APSC pluripotent cells, repairing damaged tissues and the like. Interleukin (such as IL-2 or IL-12) serves as a group of cytokines therein, and regulates an immune response by regulating and controlling the immune system. The cytokines can not only act individually to perform the function, but also can be combined with the antibody to form the antibody-cytokine fusion protein. The new protein form organically combines a unique targeting ability of the antibody with immunoregulation of the cytokines, so as to enhance an immunotherapy effect of the antibody. More importantly, the fused cytokines are transported and enriched to a tumor part through the targeting ability of the antibody, so as to effectively avoid side effects caused by individual using of the high-dose cytokines.

The term “isolated polynucleotide” refers to a polynucleotide not in a natural existing state in the nature, including a polynucleotide isolated from the nature (including the organism bodies), and also including a manually synthetic polynucleotide. The isolated polynucleotide may be a genome DNA, cDNA, mRNA or other synthetic RNAs, or a combination thereof. A plurality of nucleotide sequences used to encode the fusion protein dimer of the present invention and other polypeptide fragments are provided herein. It should be pointed out that the skilled in the art can design nucleotide sequences which are not exactly the same as the nucleotide sequences provided above but encode the same amino acid sequences according to amino acid sequences provided herein and based on codon degeneracy. These changed nucleotide sequences are also included in the scope of the present invention.

When the polynucleotide is involved, the term “vector” refers to any molecule (for example, nucleic acid, plasmid, or virus) used to transfer nucleotide encoding information into host cells. The term “expression vector” refers to a vector suitable for expressing target genes (to-be-expressed nucleotide sequences) in the host cells, and usually includes parts such as the target genes, a promoter, a terminator and a marker gene.

The term “host cell” refers to cells that have been or can be transformed through the nucleotide sequences so as to express the selected target genes. The term includes progeny of parental cells. The progeny has the selected target genes, regardless of whether the progeny is the same as an original parental cell in form or genetic constitution or not. The common host cells include bacterial, yeast and mammalian cells, for example, CHO cells.

As for a pharmaceutical composition, the used term “pharmaceutically acceptable carrier” refers to substances such as a solid or liquid diluent, a filler, an antioxidant and a stabilizer capable of being safely administered. These substances are suitable for human and/or animal drug administration without excessive adverse side effects, and meanwhile are suitable for maintaining activity of drugs or an active agent located thereinto. According to an administration route, various different carriers well known in the art may be administered, including but not limited to saccharides, starch, cellulose and derivatives thereof, maltose, gelatin, talc, calcium sulfate, vegetable oil, synthetic oil, polyols, alginic acid, a phosphatic buffer solution, an emulsifier, isotonic saline, and/or pyrogen-free water. The pharmaceutical composition provided by the present invention may be made into clinically acceptable dosage forms such as powder and injection. The pharmaceutical composition of the present invention may be administered to a subject by using any proper routes, for example, drug administration may be achieved through routes including oral, intravenous injection, intramuscular injection, subcutaneous injection, subperitoneal, rectal, sublingual, or via inhalation and transdermal.

The term “therapeutically effective amount” refers to amount of an active compound enough to cause a biological or medical reaction expected by a clinician in the body of the subject. The “therapeutically effective amount” of the fusion protein of the present invention may be determined by the skilled in the art according to factors including the administration route and the body weight, the age and the state of an illness of the subject. For example, a typical daily dosage range may be 0.01 mg-100 mg of active ingredients per kg of body weight.

The term “amino acid sequence identity” is defined as the percentage of amino acid residues in a candidate sequence that are identical to amino acid residues in a particular peptide or polypeptide sequence obtained by aligning sequences and introducing gaps when necessary to obtain the maximum percent sequence identity without considering any conservative substitutions as portion of the sequence identity. Sequence alignment can be performed in a variety of ways within the skill of the art to determine percent amino acid sequence identity, for example, using publicly available computer software, such as BLAST, BLAST-2, ALIGN, or Megalign (DNASTAR) software. Those skilled in the art can determine the appropriate parameters for measuring the alignment, including any algorithm required to obtain the maximum alignment over the entire length of the sequence being aligned.

The term “and/or” refers to elements before or after the term may exist at the same time, or only one of the elements exist. For example, “A and/or B” may be A and B, only A or only B.

The present invention improves the immunotherapy effect by utilizing advantages of immune cytokines and the monoclonal antibody. The cytokines are fused to the anti-immune checkpoint molecular antibody, such as the anti-LAG-3 antibody, the anti-PD-L1 antibody, the anti-TIGIT antibody, or the C-terminus of the heavy chain Fc region of the anti-PD-1 antibody, so that the protein antibody-cytokine fusion protein dimer aiming at anti-immune checkpoints is developed. The activity of these fusion proteins is evaluated through in-vitro activity analysis. It is found that these fusion protein dimers not only fully retain the biological activity of the antibody, but also significantly improve the biological activity of the cytokines. The cytokines mentioned in the present invention include interleukin, for example, interleukin-2 (IL-2), interleukin-12 (IL-12) and a granulocyte-macrophage colony stimulating factor (GM-CSF), where interleukin-2 and interleukin-12 participate in various links of immunoregulation, so as to enhance the immune response. More importantly, the present invention uses the target specificity of the antibody molecule, so that the immune response of immune cells is enhanced and the toxicity of the cytokines is reduced, and thus, on the premise of enhancing the efficacy of the drug, the safety of the medication is ensured.

Example 1 Design and Construction of a Monoclonal Antibody-Cytokine Fusion Protein and Expression and Purification in CHO Cells

Cytokines IL-2 or IL-12 may be connected to a heavy chain C-terminus of a monoclonal antibody through a linker sequence (Ser (Gly4Ser)3) (namely, S(G4S)3, SEQ ID NO:63), and a formed fusion protein keeps the double effects of the antibody and the cytokines. Because the functional IL-12 is constituted by two subunits of p35 and p40, IL-12 will be individually fused onto the antibody Fc in a form of the two subunits, or the two subunits of IL-12 form a single strand protein through the linker sequence (S(G4S)3) firstly, and then are connected to the C-terminus of the monoclonal antibody heavy chain through the linker sequence (S(G4S)3).

In a traditional antibody-cytokine fusion protein structure, the cytokines are connected to two heavy chains or two light chains of the antibody simultaneously so as to exist in a form of a homodimer. However, the pharmacokinetic properties of this fused immune cytokines based on IgG platform is poor. In order to fuse the cytokines into the antibody in a form of a monomer and meanwhile change the proportion of the antibody and the cytokines so as to enhance the targeting ability of the fusion protein, with an antibody heterodimer technology being adopted and based, IL-2 or IL-12 is individually fused into one heavy chain, the other chain is not connected with the cytokines so as to form an IL-12 or IL-2 monomer fusion protein, as shown in FIG. 1B and FIG. 1C respectively, or IL-2 and IL-12 are respectively connected to one heavy chain to generate the fusion protein having IL-2 and IL-12 at the same time, as shown in FIG. 1A.

The core of the antibody heterodimer technology is that the two heavy chains of the monoclonal antibody are transformed respectively to generate an asymmetric complementary structure, so that the two transformed chains can be combined to avoid generation of the homodimer. The principle of transformation is based on the following aspects: hydrophobic/spatial complementation (such as KiH and ZW1), electrostatic complementation (such as DD-KK), a spatial complementation+static electricity interaction (such as EW-RVT), spatial complementation+hydrogen bonding complementation (such as A107), and the like. In the present research, hydrophobic/spatial complementation is adopted to transform the two heavy chains of the monoclonal antibody to form the two different heavy chains, named Fm1 and Fm2 (FIG. 1) respectively, then one cytokine is connected to the C-terminus of one heavy chain, and the other heavy chain is not connected with the cytokine; or one cytokine is connected to the heavy chain Fm1, the other cytokine is connected to the heavy chain Fm2, so that the number of each cytokine contained in the finally formed heterodimer is one, instead of two in the homodimer.

The monoclonal antibody-cytokine fusion protein constructed through the above method will be expressed in CHO-3E7 cells, and then is subjected to Protein A affinity chromatography and molecular sieve purification to obtain the protein used for in-vitro analysis.

Example 2 In-Vitro Activity Analysis Experiment of a Monoclonal Antibody-Cytokine Fusion Protein

2.1: IL-2 Activity Detection:

(1) Cell culture and preparation: a piece of CTLL-2 cells is taken out of a liquid nitrogen container for recovery, and are cultured in a 5% CO2 incubator at 37° C., and the cells after recovery are subjected to secondary and enlarged culture according to a total cell count required by an activity detection experiment design.

(2) Plating: the cultured CTLL-2 cells are sampled and counted, and a moderate amount of cells are taken and inoculated into a 96-well plate.

(3) Sample dilution: the fusion protein solution sample obtained through purification and IL-2 control (100 μg/ml) are subjected to gradient dilution, each sample is provided with 9 concentration gradients, and each concentration is provided with 3 repetition holes.

(4) Sample adding: the fusion protein sample and control are added into the 96-well plate inoculated with the CTLL-2 cells, and incubated for 70 hours in the 5% CO2 incubator at 37° C.

(5) Color developing: the 96-well plate is taken out of the incubator, illuminating lamps of a biosafety cabinet are turned off, light is avoided, a Promega Substrate Cell Titer 96 Aqueous One Solution Reagent reagent is added into each experimental hole, and a reaction is performed for about 3 h at 37° C. under a light avoiding condition. After the reaction is completed, the 96-well plate is placed on a microplate reader, an OD value is detected under a 490 nm wave length.

2.2: IL-12 Activity Detection:

(1) Cell culture and preparation: a piece of NK-92 cells is taken out of a liquid nitrogen container for recovery, and are cultured in a 5% CO2 incubator at 37° C., and the cells after recovery are subjected to secondary and enlarged culture according to a total cell count required by an activity detection experiment design.

(2) Plating: the cultured NK-92 cells are sampled and counted, and a moderate amount of cells are taken and inoculated into the 96-well plate.

(3) Sample dilution: a to-be-detected fusion protein solution sample obtained through purification and IL-12 control (100 μg/ml) are dissolved and subjected to gradient dilution, each sample is provided with 9 concentration gradients, and each concentration is provided with 3 repetition holes.

(4) Sample adding: the fusion protein sample and control are added into the 96-well plate inoculated with the NK-92 cells, and incubated for 24 hours in the 5% CO2 incubator at 37° C.

(5) Sample collection: the 96-well plate is taken out of the incubator, and subjected to 4-degree centrifugation at 3700 rpm for 20 min, a supernatant is absorbed, labeling is performed, and preservation is performed at −20° C.

(6) Detection: the cryopreserved sample is taken out, and a content of human IFN gamma in the sample is detected according to operation steps of a human IFN gamma ELISA MAX™ Standard kit. The 96-well plate is placed on a microplate reader, and an OD value is detected under a 450 nm wave length.

2.3: Testing of Activity of an Anti-PD-1/PD-L1 Antibody-Cytokine Fusion Protein Through PD-1/PD-L1 Pathway Blocking Experiment

An experiment for detecting an in-vitro function of an anti-PD-1/PD-L1 McAb-cytokine fusion protein sample uses a PD-1/PD-L1 blocking function report gene kit (PD-1/PD-L1 Blockade Bioassay, a product number of a Promega kit is J1250) from Promega. The detection system of the kit is composed of two kinds of cell lines transformed by genetic engineering. A stimulating cell line is PD-L1 aAPC/CHO-K1 cells, and the cells stably express human PD-L1 and a cell surface protein capable of activating a homologous TCR in a mode independent of antigen. An effector cell line is a Jurkat T cell line, and the cell line stably expresses a luciferase reporter gene induced by human PD-1 and NFAT. When the two types of cells are co-cultured, PD-1/PD-L1 interact to inhibit TCR signal transduction and NFAT-mediated luciferase activity. Adding of the anti-PD-1/PD-L1 antibodies can block combination of PD-1 and PD-L1, so as to activate a TCR signaling pathway, enhance the NFAT-mediated luciferase activity, and generate chemiluminescence.

Firstly, the Jurkat T cells of the effector cell line are laid in the 96-well plate, and then the anti-PD-1/PD-L1 antibody-cytokine fusion protein sample and the PD-L1 aAPC/CHO-K1 cells of the stimulating cell line are added. The system is incubated for 6 hours at 37° C. Then a Bio-Glo™ fluorescent detection reagent is added, and incubation is performed for 5-10 minutes at the room temperature. Finally, a chemiluminescence signal reader is used to read fluorescence signals in the 96-well plate. The experiment uses a form of 8 concentrations and three repetition holes, a relative fluorescence unit serves as a y-axis, and the concentration of the antibody sample serves as an x-axis to draw a four-parameter curve. GraphPad Prism software is used to analyze the curve and obtain an EC₅₀ value of the anti-PD-1/PD-L1 antibody-cytokine fusion protein sample.

2.4: In-Vitro Activity Analysis of an Anti-TIGIT Antibody-Cytokine Fusion Protein

An experiment for detecting an in-vitro function of an anti-TIGIT McAb-cytokine fusion protein sample uses a TIGIT/CD155 blocking function report gene kit (TIGIT/CD155 Blockade Bioassay, a product number of a Promega kit is J2201) from Promega. A detection system of the kit is composed of two kinds of cell lines transformed by genetic engineering. A stimulating cell line is CD155 aAPC/CHO-K1 cells, and the cells express human CD-155 and a cell surface protein capable of activating a homologous TCR complex in a mode independent of antigen. An effector cell line is a Jurkat T cell line, and the cell line expresses human TIGIT and a luciferase reporter gene driven by a natural promoter and generating a response for both TCR activation and CD226 co-stimulation. When the two types of cells are co-cultured, TIGIT inhibits CD226 activation and promoter-mediated luminescence. Adding of the anti-TIGIT antibody can block interaction of TIGIT and CD155 or inhibit an ability of TIGIT for preventing CD226 from forming a homodimer, so as to recover promoter-mediated chemiluminescence.

Firstly, the Jurkat T cells of the effector cell line are laid in the 96-well plate, and then the anti-TIGIT antibody-cytokine fusion protein sample and the CD155 aAPC/CHO-K1 cells of the stimulating cell line are added. The system is incubated for 6 hours at 37° C. Then a Bio-Glo™ fluorescent detection reagent is added, and incubation is performed for 5-10 minutes at the room temperature. Finally, a chemiluminescence signal reader is used to read fluorescence signals in the 96-well plate. The experiment uses a form of 8 concentrations and three repetition holes, a relative fluorescence unit serves as a y-axis, and the concentration of the antibody sample serves as an x-axis to draw a four-parameter curve. GraphPad Prism software is used to analyze the curve and obtain an EC₅₀ value of the anti-TIGIT antibody-cytokine fusion protein sample.

2.5: In-Vitro Activity Analysis of an Anti-LAG-3 Antibody-Cytokine Fusion Protein

An experiment for detecting an in-vitro function of an anti-LAG3 McAb-cytokine fusion protein sample uses an LAG3 blocking function report gene kit (LAG3 Blockade Bioassay, a product number of a Promega kit is CS194804) from Promega. A detection system of the kit is composed of two kinds of cell lines transformed by genetic engineering. A stimulating cell line is aAPC/Raji cells, and the cells express MHC II and can activate a TCR complex. An effector cell line is a Jurkat T cell line, and the cell line expresses human LAG3 and a luciferase reporter gene driven by a natural promoter and generating a response for TCR activation. When the two types of cells are co-cultured, LAG3 inhibits MHCII activation and promoter-mediated luminescence. Adding of the anti-LAG3 antibody can block interaction of LAG3 and MHCII so as to recover promoter-mediated chemiluminescence.

Firstly, the Jurkat T cells of the effector cell line are laid in the 96-well plate, and then the anti-LAG-3 antibody-cytokine fusion protein sample and the aAPC/Raji cells of the stimulating cell line are added. The system is incubated for 6 hours at 37° C. Then a Bio-Glo™ fluorescent detection reagent is added, and incubation is performed for 5-10 minutes at the room temperature. Finally, a chemiluminescence signal reader is used to read fluorescence signals in the 96-well plate. The experiment uses a form of 8 concentrations and three repetition holes, a relative fluorescence unit serves as a y-axis, and the concentration of the antibody sample serves as an x-axis to draw a four-parameter curve. GraphPad Prism software is used to analyze the curve and obtain an EC₅₀ value of the anti-LAGS antibody-cytokine fusion protein sample.

Experiment Results and Analysis

1. Construction of an Immune Cytokine Fusion Protein Based on a Monoclonal Antibody Heterodimer

The research constructs a series of antibody-cytokine fusion proteins based on a monoclonal antibody heterodimer. The components for constructing the fusion proteins are the monoclonal antibody, cytokines IL-12 and IL-2. The present invention adopts the four different monoclonal antibodies for testing whether the technology platform is suitable for most of the monoclonal antibodies or not. The four different monoclonal antibodies are the anti-LAG-3 monoclonal antibody, the anti-PD-L1 monoclonal antibody, the anti-TIGIT monoclonal antibody and the anti-P-1 monoclonal antibody respectively. The anti-LAG-3 monoclonal antibody is Relatlimab from BMS, and belongs to a human IgG4 antibody. The anti-PD-L1 monoclonal antibody is Atezolizumab from Roche, and belongs to a human IgG1 antibody. The anti-TIGIT monoclonal antibody is Tiragolumab from Roche, and belongs to a human IgG1 antibody. The anti-PD-1 monoclonal antibody is from a patent application WO2018119474, and belongs to a human IgG4 antibody.

Firstly, encoding sequences of the heavy chain or the light chain of all the monoclonal antibodies are inserted between multiple cloning sites EcoRI and HindIII of a pTT5 expression vector, and meanwhile, a KOZAK sequence GCCGCCACC and a signal peptide sequence (SEQ ID NO:2) are further added in front of an initiation codon ATG.

In order to generate the heterodimer, a knob-into-holes (KIH) technology is utilized to transform the heavy chain Fc regions of the monoclonal antibodies, a mutation site combination of one heavy chain Fc chain is T366W/S354C, and a mutation site combination of the other heavy chain Fc chain is T366S/L368A/Y407V/Y349C. In addition, if the last amino acid of the C-terminus of the two transformed heavy chains is K, it will also be removed. DNA sequences for encoding IL-12 and IL-2 are connected to the C-terminus of the heavy chains through the linker sequence (S(G4S)3) in a Gibson assembling method respectively, so as to generate plasmids for encoding the fusion protein with the different heavy chains.

As mentioned above, the anti-LAG-3 monoclonal antibody is composed of a heavy chain H1 and a light chain L1. The heavy chain H1 is transformed by the KIH technology to generate 2 heavy chain mutants H2 and H3. The IL-12 sequence is connected to a C-terminus of the heavy chain H2 through the linker sequence (S(G4S)3) to generate a new polypeptide, called H2a. The IL-2 sequence is respectively connected to a C-terminus of the heavy chains H2 and H3 through the linker sequence (S(G4S)3) to generate new polypeptides, called H2b and H3a.

The anti-PD-L1 monoclonal antibody is composed of a heavy chain H4 and a light chain L2. The heavy chain H4 is transformed by the KIH technology to generate 2 heavy chain mutants H5 and H6. The IL-12 sequence is connected to a C-terminus of the heavy chain H5 through the linker sequence (S(G4S)3) to generate a new polypeptide, called H5a. The IL-2 sequence is respectively connected to a C-terminus of the heavy chain H5 and H6 through the linker sequence (S(G4S)3) to generate new polypeptides, called H5b and H6a.

The anti-TIGIT monoclonal antibody is composed of a heavy chain H7 and a light chain L3. The heavy chain H7 is transformed by the KIH technology to generate 2 heavy chain mutants H8 and H9. The IL-12 sequence is connected to a C-terminus of the heavy chain H8 through the linker sequence (S(G4S)3) to generate a new polypeptide, called H8a. The IL-2 sequence is respectively connected to a C-terminus of the heavy chains H8 and H9 through the linker sequence (S(G4S)3) to generate new polypeptides, called H8b and H9a.

The anti-PD-1 monoclonal antibody is composed of a heavy chain H10 and a light chain L4. The heavy chain H10 is transformed by the KIH technology to generate 2 heavy chain mutants H11 and H12. The IL-12 sequence is connected to a C-terminus of the heavy chain H11 through the linker sequence (S(G4S)3) to generate a new polypeptide, called H11a. The IL-2 sequence is respectively connected to a C-terminus of the heavy chains H11 and H12 through the linker sequence (S(G4S)3) to generate new polypeptides, called H11b and H12a.

These constructed fusion proteins are combined with non-transformed parental light chains so as to generate a series of antibody-cytokine fusion proteins. The light chain L1 and the two heavy chains H2a and H3a are combined to generate the antibody-cytokine fusion protein mLAG301, the light chain L1 and the two heavy chains H2a and H3 are combined to generate the antibody-cytokine fusion protein mLAG302, and the light chain L1 and the two heavy chains H2b and H3 are combined to generate the antibody-cytokine fusion protein mLAG303; the light chain L2 and the two heavy chains H5a and H6a are combined to generate the antibody-cytokine fusion protein mPDL101, the light chain L2 and the two heavy chains H5a and H6 are combined to generate the antibody-cytokine fusion protein mPDL102, and the light chain L2 and the two heavy chains H5b and H6 are combined to generate the antibody-cytokine fusion protein mPDL103; the light chain L3 and the two heavy chains H8a and H9a are combined to generate the antibody-cytokine fusion protein mTIGIT01, the light chain L3 and the two heavy chains H8a and H9 are combined to generate the antibody-cytokine fusion protein mTIGIT02, and the light chain L3 and the two heavy chains H8b and H9 are combined to generate the antibody-cytokine fusion protein mTIGIT03; and the light chain L4 and the two heavy chains H11a and H12a are combined to generate the antibody-cytokine fusion protein mPD101, the light chain L4 and the two heavy chains H11a and H12 are combined to generate the antibody-cytokine fusion protein mPD102, and the light chain L4 and the two heavy chains H11b and H12 are combined to generate the antibody-cytokine fusion protein mPD103. The information of the plasmids and the proteins constructed in the experiment is shown in table 1.

TABLE 1 Construction of plasmids and proteins in an antibody-cytokine fusion protein Fusion Amino acid sequence protein Component Plasmid SEQ ID NO: mLAG300 H1 pTT5-mLAG3-H 8 L1 pTT5-mLAG3-L 10 / H2 pTT5-mLAG3-H1 12 / H3 pTT5-mLAG3-H2 14 mLAG301 H2a pTT5-mLAG3a-IL12 16 H3a pTT5-mLAG3a-IL2 18 L1 pTT5-mLAG3-L 8 mLAG302 H3 pTT5-mLAG3-H2 14 H2a pTT5-mLAG3a-IL12 16 L1 pTT5-mLAG3-L 8 mLAG303 H3 pTT5-mLAG3-H2 14 H2b pTT5-mLAG3b-IL2 20 L1 pTT5-mLAG3-L 8 mPDL100 H4 pTT5-mPDL1-H 22 L2 pTT5-mPDL1-L 24 / H5 pTT5-mPDL1-H1 26 / H6 pTT5-mPDL1-H2 28 mPDL101 H5a pTT5-mTIGITa-IL12 30 H6a pTT5-mTIGITa-IL2 32 L2 pTT5-mPDL1-L 24 mPDL102 H6 pTT5-mPDL1-H2 28 H5a pTT5-mTIGITa-IL12 30 L2 pTT5-mPDL1-L 24 mPDL103 H6 pTT5-mPDL1-H2 28 H5b pTT5-mTIGITb-IL2 34 L2 pTT5-mPDL1-L 24 mTIGIT00 H7 pTT5-mTIGIT-H 36 L3 pTT5-mTIGIT-L 38 / H8 pTT5-mTIGIT-H1 40 / H9 pTT5-mTIGIT-H2 42 mTIGIT01 H8a pTT5-mTIGITa-IL12 44 H9a pTT5-mTIGITa-IL2 46 L3 pTT5-mTIGIT-L 38 mTIGIT02 H9 pTT5-mTIGIT-H2 42 H8a pTT5-mTIGITa-IL12 44 L3 pTT5-mTIGIT-L 38 mTIGIT03 H9 pTT5-mTIGIT-H2 42 H8b pTT5-mTIGITb-IL2 48 L3 pTT5-mTIGIT-L 38 mPD100 H10 pTT5-mPD1-H 50 L4 pTT5-mPD1-L 52 / H11 pTT5-mPD1-H1 54 / H12 pTT5-mPD1-H2 56 mPD101 H11a pTT5-mPD1a-IL12 58 H12a pTT5-mPD1a-IL2 60 L4 pTT5-mPD1-L 52 mPD102 H12 pTT5-mPD1-H2 56 H11a pTT5-mPD1a-IL12 58 L4 pTT5-mPD1-L 52 mPD103 H12 pTT5-mPD1-H2 56 H11b pTT5-mPD1b-IL2 62 L4 pTT5-mPD1-L 52

The above fusion protein plasmids constructed on the pTT5 expression vectors transiently transfect CHO-3E7 cells through a PEI transfection reagent, and then culturing is performed at 37° C. for 6 days. A culture solution supernatant is centrifugally taken, the fusion proteins are first purified through Protein A affinity columns, then a further purification is performed through a molecular sieve, and the final purity reaches 95% or above.

2. Result Analysis of Cytokine Activity Detection

In order to prove broad spectrum of the technological platform of the monoclonal antibody-cytokine fusion protein based on the antibody heterodimer technology, the present invention utilizes the 4 different antibodies to construct the series of antibody-cytokine fusion proteins. As for the antibody-cytokine fusion protein fused with IL-2, compared with biological activity of free IL-2, the activity of IL-2 of all the fusion proteins is obviously enhanced (FIGS. 2, 3, 4, and 5), and this shows that a structure of the monoclonal antibody-cytokine fusion protein based on the antibody heterodimer technology can significantly improve the activity of IL-2.

As for the fusion protein fused with IL-12, compared with biological activity of free IL-12, the activity of IL-12 of the antibody-cytokine fusion proteins constructed by utilizing the 4 different antibodies is enhanced or unchanged (FIGS. 6, 7, 8 and 9). The activity of IL-12 of the fusion protein only fused with IL-2 is kept unchanged, and this shows that the structure of the monoclonal antibody-cytokine fusion protein based on the antibody heterodimer technology can well maintain the activity of IL-12. More importantly, the IL-12 cytokine activity of the antibody-cytokine fusion protein simultaneously fused with IL-12 and IL-2 is higher than the fusion protein only fused with IL-12, this shows that IL-12 and IL-2 may have a synergistic effect, and it is further proved that the structure of the monoclonal antibody-cytokine fusion protein based on the antibody heterodimer technology can significantly improve the activity of IL-12.

3. Result Analysis of Antibody Activity Detection

In the present invention, the series of antibody-cytokine fusion proteins are constructed based on the 4 different monoclonal antibodies, and then the antibody activity in each fusion protein is detected by utilizing the antibody activity detection kit of Promega. Results show that compared with untransformed anti-LAG-3 monoclonal antibody control mLAG300, the antibody activity of the constructed heterodimer fusion protein (mLAG303) is almost the same as the control (FIG. 10), and this shows that the antibody-cytokine fusion protein based on the antibody heterodimer technology does not influence exertion of the antibody activity of the fusion protein. More importantly, the antibody activity of the heterodimer fusion protein (mLAG301 and mLAG302) fused with IL-12 is enhanced by 2 times or more than the antibody activity of the monoclonal antibody control (FIG. 10), and this shows that the antibody-cytokine fusion proteins based on the antibody heterodimer technology and fused with IL-12 can significantly enhance the antibody activity of the fusion proteins. Similarly, the antibody activity of the antibody-cytokine fusion protein transformed from the other three monoclonal antibodies (the anti-PD-L1 antibody, the anti-TIGIT antibody, and the anti-PD-1 antibody) is similar to the activity of the respective monoclonal antibody control (mPDL100, mTIGIT00 and mPD100). This shows that the antibody-cytokine fusion proteins based on the antibody heterodimer technology do not influence exertion of the antibody activity, and even enhance the antibody activity of certain fusion proteins.

In conclusion, the present invention develops the novel technological platform of the antibody-cytokine fusion proteins based on the antibody heterodimer technology, constructs the series of antibody-cytokine fusion proteins by utilizing the 4 different antibodies, and verifies that the technological platform can enhance the antibody activity or cytokine activity through a series of antibody and cytokine activity analysis experiments.

A person skilled in the art to which the present invention belongs should understand that the above described methods and materials are only exemplary, and should not be regarded as limitation to the scope of the present invention.

Sequence information DNA sequence of signal peptide (SEQ ID NO: 1): ATGGGATGGTCTTGGATCCTGCTGTTCCTGCTGAGCGTGACAGCTGGCGTGCA CTCT Amino acid sequence of signal peptide (SEQ ID NO: 2) MGWSWILLFLLSVTAGVHS DNA sequence of cytokine IL-12 (SEQ ID NO: 3) ATTTGGGAGCTGAAGAAAGACGTGTACGTGGTCGAGCTGGACTGGTACCCTGATG CCCCAGGCGAGATGGTCGTGCTGACCTGCGATACACCAGAGGAAGATGGTATCAC CTGGACACTGGATCAGTCCTCAGAGGTGCTGGGCTCTGGTAAAACACTGACCATT CAGGTGAAGGAGTTCGGTGACGCTGGACAGTACACTTGTCATAAGGGCGGGGAG GTGCTGTCTCACTCCCTGCTGCTGCTGCATAAGAAGGAGGATGGAATCTGGTCCA CTGACATCCTGAAAGACCAGAAGGAGCCAAAGAACAAAACCTTCCTGCGATGCG AGGCTAAGAACTACAGCGGCCGCTTTACATGCTGGTGGCTGACAACCATCAGCAC CGATCTGACCTTTAGCGTGAAGTCATCCAGGGGCAGTTCAGACCCTCAGGGAGTC ACATGTGGCGCCGCAACCCTGTCAGCAGAGCGAGTGCGGGGAGACAATAAGGAA TACGAGTACAGCGTCGAGTGTCAGGAGGATTCCGCATGTCCAGCTGCAGAAGAAT CCCTGCCTATCGAAGTCATGGTGGACGCTGTGCATAAACTGAAGTACGAGAATTAC ACCAGCAGCTTTTTCATCCGGGACATCATCAAGCCCGATCCACCTAAGAATCTGCA GCTGAAGCCTCTGAAAAATAGCCGACAGGTCGAAGTGTCATGGGAATACCCAGAC ACCTGGTCAACACCACACTCCTACTTCTCCCTGACCTTCTGTGTGCAGGTCCAGG GAAAAAGCAAGCGGGAAAAGAAAGATCGGGTGTTCACCGACAAGACCAGTGCTA CAGTGATTTGCCGGAAGAATGCCAGCATTTCTGTCAGAGCTCAGGACCGGTACTAT AGCTCTTCCTGGAGCGAGTGGGCTTCAGTGCCATGTTCTGGaGGCGGtGGATCTGG CGGAGGTGGAAGCGGAGGCGGTGGATCTAGAAACCTGCCCGTCGCAACCCCTGA TCCAGGGATGTTCCCCTGTCTGCATCACAGCCAGAATCTGCTGAGGGCTGTCTCCA ACATGCTGCAGAAGGCTCGACAGACCCTGGAGTTCTACCCATGTACCAGCGAAGA GATCGACCACGAGGATATCACAAAGGATAAAACCAGCACAGTGGAAGCATGCCTG CCTCTGGAACTGACCAAGAATGAGAGCTGCCTGAATAGCAGGGAGACCTCCTTCA TCACCAACGGCTCATGCCTGGCTTCAAGGAAGACCAGCTTCATGATGGCTCTGTG TCTGAGCTCTATCTATGAGGACCTGAAGATGTACCAGGTGGAGTTCAAGACCATGA ACGCCAAGCTGCTGATGGATCCAAAGAGGCAGATCTTCCTGGATCAGAATATGCT GGCAGTGATCGATGAGCTGATGCAGGCCCTGAATTTTAACAGTGAGACAGTGCCT CAGAAGAGCTCTCTGGAAGAGCCAGACTTTTACAAAACTAAGATCAAGCTGTGCA TTCTGCTGCACGCTTTCCGCATCAGAGCTGTCACTATCGATAGAGTGATGAGCTAT CTGAATGCCTCA Amino acid sequence of cytokine IL-12 full-length (SEQ ID NO: 4) IWELKKDVYVVELDWYPDAPGEMVVLTCDTPEEDGITWTLDQSSEVLGSGKTLT IQVKEFGDAGQYTCHKGGEVLSHSLLLLHKKEDGIWSTDILKDQKEPKNKTFLRCEA KNYSGRFTCWWLTTISTDLTFSVKSSRGSSDPQGVTCGAATLSAERVRGDNKEYEYS VECQEDSACPAAEESLPIEVMVDAVHKLKYENYTSSFFIRDIIKPDPPKNLQLKPLKNS RQVEVSWEYPDTWSTPHSYFSLTFCVQVQGKSKREKKDRVFTDKTSATVICRKNASI SVRAQDRYYSSSWSEWASVPCSGGGGSGGGGSGGGGSRNLPVATPDPGMFPCLHHS QNLLRAVSNMLQKARQTLEFYPCTSEEIDHEDITKDKTSTVEACLPLELTKNESCLNS RETSFITNGSCLASRKTSFMMALCLSSIYEDLKMYQVEFKTMNAKLLMDPKRQIFLD QNMLAVIDELMQALNFNSETVPQKSSLEEPDFYKTKIKLCILLHAFRIRAVTIDRVMSY LNAS DNA sequence of cytokine IL-2 (SEQ ID NO: 5) GCACCTACTTCAAGTTCTACAAAGAAAACACAGCTACAACTGGAGCATTTACT GCTGGATTTACAGATGATTTTGAATGGAATTAATAATTACAAGAATCCCAAACTCAC CAGGATGCTCACATTTAAGTTTTACATGCCCAAGAAGGCCACAGAACTGAAACAT CTTCAGTGTCTAGAAGAAGAACTCAAACCTCTGGAGGAAGTGCTAAATTTAGCTC AAAGCAAAAACTTTCACTTAAGACCCAGGGACTTAATCAGCAATATCAACGTAATA GTTCTGGAACTAAAGGGATCTGAAACAACATTCATGTGTGAATATGCTGATGAGAC AGCAACCATTGTAGAATTTCTGAACAGATGGATTACCTTTTGTCAAAGCATCATCT CAACACTGACT Amino acid sequence of cytokine IL-2 (SEQ ID NO: 6) APTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTFKFYMPKKATELKH LQCLEEELKPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATI VEFLNRWITFCQSIISTLT DNA sequence of polypeptide chain H1 (SEQ ID NO: 7) CAAGTGCAGCTGCAGCAGTGGGGCGCTGGCCTGCTGAAGCCTTCCGAGACAC TGTCTCTGACCTGTGCTGTTTACGGCGGCTCCTTCTCCGACTACTACTGGAACTGG ATCAGACAGCCTCCTGGCAAGGGCCTGGAATGGATCGGCGAGATCAACCACAGA GGCTCTACCAACTCCAACCCATCTCTGAAGTCCAGAGTGACACTGAGTCTGGACA CCTCTAAGAATCAGTTCTCCCTGAAGCTGAGATCTGTGACCGCCGCCGACACCGC TGTGTACTACTGCGCCTTCGGCTACTCCGACTATGAATACAACTGGTTCGACCCTT GGGGCCAGGGCACCCTGGTGACCGTGAGCTCTGCTTCCACAAAAGGCCCTAGCG TGTTTCCTCTGGCCCCTTGCTCCAGATCCACTTCTGAGTCCACCGCTGCTCTCGGC TGCCTGGTGAAGGACTACTTCCCCGAACCCGTCACCGTGTCCTGGAACAGCGGCG CCTTGACCTCCGGCGTGCACACCTTCCCTGCCGTGCTGCAGAGTAGCGGACTGTAT AGCCTGTCCAGCGTCGTGACCGTGCCTTCCTCTTCCCTGGGCACAAAGACATACA CCTGCAACGTGGACCACAAGCCCTCTAACACCAAAGTGGATAAGCGGGTAGAGTC CAAGTACGGCCCCCCTTGCCCTCCTTGTCCTGCTCCTGAGTTCCTGGGAGGCCCTT CTGTGTTTCTGTTCCCCCCCAAACCTAAGGATACCCTGATGATCTCTCGGACCCCT GAGGTGACCTGCGTGGTGGTGGACGTGTCACAAGAGGATCCTGAAGTGCAGTTTA ATTGGTACGTGGACGGCGTGGAAGTGCATAACGCCAAAACCAAACCACGCGAAG AACAGTTCAACTCCACCTACAGAGTCGTCAGCGTGCTGACCGTGCTGCACCAGGA TTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTGTCCAACAAGGGACTGCCTTC TTCCATCGAGAAGACCATCTCCAAGGCCAAGGGCCAGCCTCGGGAGCCTCAGGT GTACACCCTCCCTCCATCCCAGGAAGAGATGACCAAGAACCAGGTGTCCCTGACC TGTCTGGTCAAGGGGTTCTACCCCTCTGACATCGCCGTGGAGTGGGAGTCTAATG GCCAACCCGAGAACAACTACAAGACCACCCCTCCAGTGCTGGACTCTGATGGATC TTTCTTCCTGTACTCCCGGCTGACAGTGGACAAGAGCAGATGGCAGGAAGGCAAC GTGTTCTCCTGCTCCGTGATGCACGAGGCCCTGCATAACCACTATACCCAGAAGTC TCTGTCTCTGTCTCTGGGA Amino acid sequence of polypeptide chain H1 (SEQ ID NO: 8) QVQLQQWGAGLLKPSETLSLTCAVYGGSFSDYYWNWIRQPPGKGLEWIGEINHR GSTNSNPSLKSRVTLSLDTSKNQFSLKLRSVTAADTAVYYCAFGYSDYEYNWFDPWG QGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSG VHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCP PCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEV HNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKG QPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVL DSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG DNA sequence of polypeptide chain L1 (SEQ ID NO: 9) GAGATCGTGCTGACCCAGTCCCCCGCCACTTTGTCTCTGTCTCCTGGCGAGAG AGCTACCCTGTCCTGCAGAGCCTCTCAGTCCATCTCTTCCTACCTGGCCTGGTACC AGCAGAAACCTGGACAGGCTCCTCGGCTGCTGATCTACGACGCTTCTAATCGCGC TACAGGCATCCCTGCTAGATTCTCCGGCTCTGGCTCCGGCACCGATTTTACCCTGA CCATCTCCAGCCTCGAGCCCGAAGATTTTGCCGTGTACTACTGCCAGCAGCGGTCC AACTGGCCTCTGACCTTCGGCCAGGGAACAAACCTGGAAATCAAGCGGACCGTG GCCGCCCCTAGCGTCTTCATCTTCCCTCCTTCCGACGAACAACTGAAGTCTGGCAC CGCTAGCGTGGTGTGCCTGCTGAACAACTTCTACCCCAGAGAGGCCAAAGTGCAG TGGAAGGTGGACAACGCCCTGCAGTCTGGCAACTCCCAAGAGTCCGTGACCGAG CAGGACTCCAAGGACTCTACCTATTCCCTGTCTTCTACACTGACCCTGAGCAAGGC CGACTACGAGAAGCACAAGGTGTACGCCTGCGAGGTGACCCACCAGGGCCTGTC CAGCCCAGTCACCAAGTCCTTCAACAGAGGCGAATGT Amino acid sequence of polypeptide chain L1 (SEQ ID NO: 10) EIVLTQSPATLSLSPGERATLSCRASQSISSYLAWYQQKPGQAPRLLIYDASNRATG IPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPLTFGQGTNLEIKRTVAAPSVFIF PPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYS LSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC DNA sequence of polypeptide chain H2 (SEQ ID NO: 11) CAAGTGCAGCTGCAGCAGTGGGGCGCTGGCCTGCTGAAGCCTTCCGAGACAC TGTCTCTGACCTGTGCTGTTTACGGCGGCTCCTTCTCCGACTACTACTGGAACTGG ATCAGACAGCCTCCTGGCAAGGGCCTGGAATGGATCGGCGAGATCAACCACAGA GGCTCTACCAACTCCAACCCATCTCTGAAGTCCAGAGTGACACTGAGTCTGGACA CCTCTAAGAATCAGTTCTCCCTGAAGCTGAGATCTGTGACCGCCGCCGACACCGC TGTGTACTACTGCGCCTTCGGCTACTCCGACTATGAATACAACTGGTTCGACCCTT GGGGCCAGGGCACCCTGGTGACCGTGAGCTCTGCTTCCACAAAAGGCCCTAGCG TGTTTCCTCTGGCCCCTTGCTCCAGATCCACTTCTGAGTCCACCGCTGCTCTCGGC TGCCTGGTGAAGGACTACTTCCCCGAACCCGTCACCGTGTCCTGGAACAGCGGCG CCTTGACCTCCGGCGTGCACACCTTCCCTGCCGTGCTGCAGAGTAGCGGACTGTAT AGCCTGTCCAGCGTCGTGACCGTGCCTTCCTCTTCCCTGGGCACAAAGACATACA CCTGCAACGTGGACCACAAGCCCTCTAACACCAAAGTGGATAAGCGGGTAGAGTC CAAGTACGGCCCCCCTTGCCCTCCTTGTCCTGCTCCTGAGTTCCTGGGAGGCCCTT CTGTGTTTCTGTTCCCCCCCAAACCTAAGGATACCCTGATGATCTCTCGGACCCCT GAGGTGACCTGCGTGGTGGTGGACGTGTCACAAGAGGATCCTGAAGTGCAGTTTA ATTGGTACGTGGACGGCGTGGAAGTGCATAACGCCAAAACCAAACCACGCGAAG AACAGTTCAACTCCACCTACAGAGTCGTCAGCGTGCTGACCGTGCTGCACCAGGA TTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTGTCCAACAAGGGACTGCCTTC TTCCATCGAGAAGACCATCTCCAAGGCCAAGGGCCAGCCTCGGGAGCCTCAGGT GTACACCCTCCCTCCATGCCAGGAAGAGATGACCAAGAACCAGGTGTCCCTGTGG TGTCTGGTCAAGGGGTTCTACCCCTCTGACATCGCCGTGGAGTGGGAGTCTAATG GCCAACCCGAGAACAACTACAAGACCACCCCTCCAGTGCTGGACTCTGATGGATC TTTCTTCCTGTACTCCCGGCTGACAGTGGACAAGAGCAGATGGCAGGAAGGCAAC GTGTTCTCCTGCTCCGTGATGCACGAGGCCCTGCATAACCACTATACCCAGAAGTC TCTGTCTCTGTCTCTGGGA Amino acid sequence of polypeptide chain H2 (SEQ ID NO: 12) QVQLQQWGAGLLKPSETLSLTCAVYGGSFSDYYWNWIRQPPGKGLEWIGEINHR GSTNSNPSLKSRVTLSLDTSKNQFSLKLRSVTAADTAVYYCAFGYSDYEYNWFDPWG QGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSG VHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCP PCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEV HNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKG QPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVL DSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG DNA sequence of polypeptide chain H3 (SEQ ID NO: 13) CAAGTGCAGCTGCAGCAGTGGGGCGCTGGCCTGCTGAAGCCTTCCGAGACAC TGTCTCTGACCTGTGCTGTTTACGGCGGCTCCTTCTCCGACTACTACTGGAACTGG ATCAGACAGCCTCCTGGCAAGGGCCTGGAATGGATCGGCGAGATCAACCACAGA GGCTCTACCAACTCCAACCCATCTCTGAAGTCCAGAGTGACACTGAGTCTGGACA CCTCTAAGAATCAGTTCTCCCTGAAGCTGAGATCTGTGACCGCCGCCGACACCGC TGTGTACTACTGCGCCTTCGGCTACTCCGACTATGAATACAACTGGTTCGACCCTT GGGGCCAGGGCACCCTGGTGACCGTGAGCTCTGCTTCCACAAAAGGCCCTAGCG TGTTTCCTCTGGCCCCTTGCTCCAGATCCACTTCTGAGTCCACCGCTGCTCTCGGC TGCCTGGTGAAGGACTACTTCCCCGAACCCGTCACCGTGTCCTGGAACAGCGGCG CCTTGACCTCCGGCGTGCACACCTTCCCTGCCGTGCTGCAGAGTAGCGGACTGTAT AGCCTGTCCAGCGTCGTGACCGTGCCTTCCTCTTCCCTGGGCACAAAGACATACA CCTGCAACGTGGACCACAAGCCCTCTAACACCAAAGTGGATAAGCGGGTAGAGTC CAAGTACGGCCCCCCTTGCCCTCCTTGTCCTGCTCCTGAGTTCCTGGGAGGCCCTT CTGTGTTTCTGTTCCCCCCCAAACCTAAGGATACCCTGATGATCTCTCGGACCCCT GAGGTGACCTGCGTGGTGGTGGACGTGTCACAAGAGGATCCTGAAGTGCAGTTTA ATTGGTACGTGGACGGCGTGGAAGTGCATAACGCCAAAACCAAACCACGCGAAG AACAGTTCAACTCCACCTACAGAGTCGTCAGCGTGCTGACCGTGCTGCACCAGGA TTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTGTCCAACAAGGGACTGCCTTC TTCCATCGAGAAGACCATCTCCAAGGCCAAGGGCCAGCCTCGGGAGCCTCAGGT GTGTACCCTCCCTCCATCCCAGGAAGAGATGACCAAGAACCAGGTGTCCCTGTCC TGTGCTGTCAAGGGGTTCTACCCCTCTGACATCGCCGTGGAGTGGGAGTCTAATG GCCAACCCGAGAACAACTACAAGACCACCCCTCCAGTGCTGGACTCTGATGGATC TTTCTTCCTGGTGTCCCGGCTGACAGTGGACAAGAGCAGATGGCAGGAAGGCAA CGTGTTCTCCTGCTCCGTGATGCACGAGGCCCTGCATAACCACTATACCCAGAAGT CTCTGTCTCTGTCTCTGGGA Amino acid sequence of polypeptide chain H3 (SEQ ID NO: 14) QVQLQQWGAGLLKPSETLSLTCAVYGGSFSDYYWNWIRQPPGKGLEWIGEINHR GSTNSNPSLKSRVTLSLDTSKNQFSLKLRSVTAADTAVYYCAFGYSDYEYNWFDPWG QGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSG VHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCP PCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEV HNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKG QPREPQVCTLPPSQEEMTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVL DSDGSFFLVSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG DNA sequence of polypeptide chain H2a (SEQ ID NO: 15) CAAGTGCAGCTGCAGCAGTGGGGCGCTGGCCTGCTGAAGCCTTCCGAGACAC TGTCTCTGACCTGTGCTGTTTACGGCGGCTCCTTCTCCGACTACTACTGGAACTGG ATCAGACAGCCTCCTGGCAAGGGCCTGGAATGGATCGGCGAGATCAACCACAGA GGCTCTACCAACTCCAACCCATCTCTGAAGTCCAGAGTGACACTGAGTCTGGACA CCTCTAAGAATCAGTTCTCCCTGAAGCTGAGATCTGTGACCGCCGCCGACACCGC TGTGTACTACTGCGCCTTCGGCTACTCCGACTATGAATACAACTGGTTCGACCCTT GGGGCCAGGGCACCCTGGTGACCGTGAGCTCTGCTTCCACAAAAGGCCCTAGCG TGTTTCCTCTGGCCCCTTGCTCCAGATCCACTTCTGAGTCCACCGCTGCTCTCGGC TGCCTGGTGAAGGACTACTTCCCCGAACCCGTCACCGTGTCCTGGAACAGCGGCG CCTTGACCTCCGGCGTGCACACCTTCCCTGCCGTGCTGCAGAGTAGCGGACTGTAT AGCCTGTCCAGCGTCGTGACCGTGCCTTCCTCTTCCCTGGGCACAAAGACATACA CCTGCAACGTGGACCACAAGCCCTCTAACACCAAAGTGGATAAGCGGGTAGAGTC CAAGTACGGCCCCCCTTGCCCTCCTTGTCCTGCTCCTGAGTTCCTGGGAGGCCCTT CTGTGTTTCTGTTCCCCCCCAAACCTAAGGATACCCTGATGATCTCTCGGACCCCT GAGGTGACCTGCGTGGTGGTGGACGTGTCACAAGAGGATCCTGAAGTGCAGTTTA ATTGGTACGTGGACGGCGTGGAAGTGCATAACGCCAAAACCAAACCACGCGAAG AACAGTTCAACTCCACCTACAGAGTCGTCAGCGTGCTGACCGTGCTGCACCAGGA TTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTGTCCAACAAGGGACTGCCTTC TTCCATCGAGAAGACCATCTCCAAGGCCAAGGGCCAGCCTCGGGAGCCTCAGGT GTACACCCTCCCTCCATGCCAGGAAGAGATGACCAAGAACCAGGTGTCCCTGTGG TGTCTGGTCAAGGGGTTCTACCCCTCTGACATCGCCGTGGAGTGGGAGTCTAATG GCCAACCCGAGAACAACTACAAGACCACCCCTCCAGTGCTGGACTCTGATGGATC TTTCTTCCTGTACTCCCGGCTGACAGTGGACAAGAGCAGATGGCAGGAAGGCAAC GTGTTCTCCTGCTCCGTGATGCACGAGGCCCTGCATAACCACTATACCCAGAAGTC TCTGTCTCTGTCTCTGGGATCAGGTGGAGGCGGTAGTGGCGGAGGCGGTTCAGGC GGAGGCGGATCTATTTGGGAGCTGAAGAAAGACGTGTACGTGGTCGAGCTGGACT GGTACCCTGATGCCCCAGGCGAGATGGTCGTGCTGACCTGCGATACACCAGAGGA AGATGGTATCACCTGGACACTGGATCAGTCCTCAGAGGTGCTGGGCTCTGGTAAA ACACTGACCATTCAGGTGAAGGAGTTCGGTGACGCTGGACAGTACACTTGTCATA AGGGCGGGGAGGTGCTGTCTCACTCCCTGCTGCTGCTGCATAAGAAGGAGGATGG AATCTGGTCCACTGACATCCTGAAAGACCAGAAGGAGCCAAAGAACAAAACCTT CCTGCGATGCGAGGCTAAGAACTACAGCGGCCGCTTTACATGCTGGTGGCTGACA ACCATCAGCACCGATCTGACCTTTAGCGTGAAGTCATCCAGGGGCAGTTCAGACC CTCAGGGAGTCACATGTGGCGCCGCAACCCTGTCAGCAGAGCGAGTGCGGGGAG ACAATAAGGAATACGAGTACAGCGTCGAGTGTCAGGAGGATTCCGCATGTCCAGC TGCAGAAGAATCCCTGCCTATCGAAGTCATGGTGGACGCTGTGCATAAACTGAAG TACGAGAATTACACCAGCAGCTTTTTCATCCGGGACATCATCAAGCCCGATCCACC TAAGAATCTGCAGCTGAAGCCTCTGAAAAATAGCCGACAGGTCGAAGTGTCATGG GAATACCCAGACACCTGGTCAACACCACACTCCTACTTCTCCCTGACCTTCTGTGT GCAGGTCCAGGGAAAAAGCAAGCGGGAAAAGAAAGATCGGGTGTTCACCGACA AGACCAGTGCTACAGTGATTTGCCGGAAGAATGCCAGCATTTCTGTCAGAGCTCA GGACCGGTACTATAGCTCTTCCTGGAGCGAGTGGGCTTCAGTGCCATGTTCTGGAG GCGGTGGATCTGGCGGAGGTGGAAGCGGAGGCGGTGGATCTAGAAACCTGCCCG TCGCAACCCCTGATCCAGGGATGTTCCCCTGTCTGCATCACAGCCAGAATCTGCTG AGGGCTGTCTCCAACATGCTGCAGAAGGCTCGACAGACCCTGGAGTTCTACCCAT GTACCAGCGAAGAGATCGACCACGAGGATATCACAAAGGATAAAACCAGCACAG TGGAAGCATGCCTGCCTCTGGAACTGACCAAGAATGAGAGCTGCCTGAATAGCAG GGAGACCTCCTTCATCACCAACGGCTCATGCCTGGCTTCAAGGAAGACCAGCTTC ATGATGGCTCTGTGTCTGAGCTCTATCTATGAGGACCTGAAGATGTACCAGGTGGA GTTCAAGACCATGAACGCCAAGCTGCTGATGGATCCAAAGAGGCAGATCTTCCTG GATCAGAATATGCTGGCAGTGATCGATGAGCTGATGCAGGCCCTGAATTTTAACAG TGAGACAGTGCCTCAGAAGAGCTCTCTGGAAGAGCCAGACTTTTACAAAACTAA GATCAAGCTGTGCATTCTGCTGCACGCTTTCCGCATCAGAGCTGTCACTATCGATA GAGTGATGAGCTATCTGAATGCCTCA Amino acid sequence of polypeptide chain H2a (SEQ ID NO: 16) QVQLQQWGAGLLKPSETLSLTCAVYGGSFSDYYWNWIRQPPGKGLEWIGEINHR GSTNSNPSLKSRVTLSLDTSKNQFSLKLRSVTAADTAVYYCAFGYSDYEYNWFDPWG QGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSG VHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCP PCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEV HNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKG QPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVL DSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGSGGGGSGG GGSGGGGSIWELKKDVYVVELDWYPDAPGEMVVLTCDTPEEDGITWTLDQSSEVLG SGKTLTIQVKEFGDAGQYTCHKGGEVLSHSLLLLHKKEDGIWSTDILKDQKEPKNKT FLRCEAKNYSGRFTCWWLTTISTDLTFSVKSSRGSSDPQGVTCGAATLSAERVRGDN KEYEYSVECQEDSACPAAEESLPIEVMVDAVHKLKYENYTSSFFIRDIIKPDPPKNLQL KPLKNSRQVEVSWEYPDTWSTPHSYFSLTFCVQVQGKSKREKKDRVFTDKTSATVIC RKNASISVRAQDRYYSSSWSEWASVPCSGGGGSGGGGSGGGGSRNLPVATPDPGMFP CLHHSQNLLRAVSNMLQKARQTLEFYPCTSEEIDHEDITKDKTSTVEACLPLELTKNE SCLNSRETSFITNGSCLASRKTSFMMALCLSSIYEDLKMYQVEFKTMNAKLLMDPKR QIFLDQNMLAVIDELMQALNFNSETVPQKSSLEEPDFYKTKIKLCILLHAFRIRAVTIDR VMSYLNAS DNA sequence of polypeptide chain H3a (SEQ ID NO: 17) CAAGTGCAGCTGCAGCAGTGGGGCGCTGGCCTGCTGAAGCCTTCCGAGACAC TGTCTCTGACCTGTGCTGTTTACGGCGGCTCCTTCTCCGACTACTACTGGAACTGG ATCAGACAGCCTCCTGGCAAGGGCCTGGAATGGATCGGCGAGATCAACCACAGA GGCTCTACCAACTCCAACCCATCTCTGAAGTCCAGAGTGACACTGAGTCTGGACA CCTCTAAGAATCAGTTCTCCCTGAAGCTGAGATCTGTGACCGCCGCCGACACCGC TGTGTACTACTGCGCCTTCGGCTACTCCGACTATGAATACAACTGGTTCGACCCTT GGGGCCAGGGCACCCTGGTGACCGTGAGCTCTGCTTCCACAAAAGGCCCTAGCG TGTTTCCTCTGGCCCCTTGCTCCAGATCCACTTCTGAGTCCACCGCTGCTCTCGGC TGCCTGGTGAAGGACTACTTCCCCGAACCCGTCACCGTGTCCTGGAACAGCGGCG CCTTGACCTCCGGCGTGCACACCTTCCCTGCCGTGCTGCAGAGTAGCGGACTGTAT AGCCTGTCCAGCGTCGTGACCGTGCCTTCCTCTTCCCTGGGCACAAAGACATACA CCTGCAACGTGGACCACAAGCCCTCTAACACCAAAGTGGATAAGCGGGTAGAGTC CAAGTACGGCCCCCCTTGCCCTCCTTGTCCTGCTCCTGAGTTCCTGGGAGGCCCTT CTGTGTTTCTGTTCCCCCCCAAACCTAAGGATACCCTGATGATCTCTCGGACCCCT GAGGTGACCTGCGTGGTGGTGGACGTGTCACAAGAGGATCCTGAAGTGCAGTTTA ATTGGTACGTGGACGGCGTGGAAGTGCATAACGCCAAAACCAAACCACGCGAAG AACAGTTCAACTCCACCTACAGAGTCGTCAGCGTGCTGACCGTGCTGCACCAGGA TTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTGTCCAACAAGGGACTGCCTTC TTCCATCGAGAAGACCATCTCCAAGGCCAAGGGCCAGCCTCGGGAGCCTCAGGT GTGTACCCTCCCTCCATCCCAGGAAGAGATGACCAAGAACCAGGTGTCCCTGTCC TGTGCTGTCAAGGGGTTCTACCCCTCTGACATCGCCGTGGAGTGGGAGTCTAATG GCCAACCCGAGAACAACTACAAGACCACCCCTCCAGTGCTGGACTCTGATGGATC TTTCTTCCTGGTGTCCCGGCTGACAGTGGACAAGAGCAGATGGCAGGAAGGCAA CGTGTTCTCCTGCTCCGTGATGCACGAGGCCCTGCATAACCACTATACCCAGAAGT CTCTGTCTCTGTCTCTGGGATCAGGTGGAGGCGGTAGTGGCGGAGGCGGTTCAGG CGGAGGCGGATCTGCACCTACTTCAAGTTCTACAAAGAAAACACAGCTACAACTG GAGCATTTACTGCTGGATTTACAGATGATTTTGAATGGAATTAATAATTACAAGAAT CCCAAACTCACCAGGATGCTCACATTTAAGTTTTACATGCCCAAGAAGGCCACAG AACTGAAACATCTTCAGTGTCTAGAAGAAGAACTCAAACCTCTGGAGGAAGTGCT AAATTTAGCTCAAAGCAAAAACTTTCACTTAAGACCCAGGGACTTAATCAGCAATA TCAACGTAATAGTTCTGGAACTAAAGGGATCTGAAACAACATTCATGTGTGAATAT GCTGATGAGACAGCAACCATTGTAGAATTTCTGAACAGATGGATTACCTTTTGTCA AAGCATCATCTCAACACTGACT Amino acid sequence of polypeptide chain H3a (SEQ ID NO: 18) QVQLQQWGAGLLKPSETLSLTCAVYGGSFSDYYWNWIRQPPGKGLEWIGEINHR GSTNSNPSLKSRVTLSLDTSKNQFSLKLRSVTAADTAVYYCAFGYSDYEYNWFDPWG QGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSG VHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCP PCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEV HNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKG QPREPQVCTLPPSQEEMTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVL DSDGSFFLVSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGSGGGGSGG GGSGGGGSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTFKFYMPKK ATELKHLQCLEEELKPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYA DETATIVEFLNRWITFCQSIISTLT DNA sequence of polypeptide chain H2b (SEQ ID NO: 19) CAAGTGCAGCTGCAGCAGTGGGGCGCTGGCCTGCTGAAGCCTTCCGAGACAC TGTCTCTGACCTGTGCTGTTTACGGCGGCTCCTTCTCCGACTACTACTGGAACTGG ATCAGACAGCCTCCTGGCAAGGGCCTGGAATGGATCGGCGAGATCAACCACAGA GGCTCTACCAACTCCAACCCATCTCTGAAGTCCAGAGTGACACTGAGTCTGGACA CCTCTAAGAATCAGTTCTCCCTGAAGCTGAGATCTGTGACCGCCGCCGACACCGC TGTGTACTACTGCGCCTTCGGCTACTCCGACTATGAATACAACTGGTTCGACCCTT GGGGCCAGGGCACCCTGGTGACCGTGAGCTCTGCTTCCACAAAAGGCCCTAGCG TGTTTCCTCTGGCCCCTTGCTCCAGATCCACTTCTGAGTCCACCGCTGCTCTCGGC TGCCTGGTGAAGGACTACTTCCCCGAACCCGTCACCGTGTCCTGGAACAGCGGCG CCTTGACCTCCGGCGTGCACACCTTCCCTGCCGTGCTGCAGAGTAGCGGACTGTAT AGCCTGTCCAGCGTCGTGACCGTGCCTTCCTCTTCCCTGGGCACAAAGACATACA CCTGCAACGTGGACCACAAGCCCTCTAACACCAAAGTGGATAAGCGGGTAGAGTC CAAGTACGGCCCCCCTTGCCCTCCTTGTCCTGCTCCTGAGTTCCTGGGAGGCCCTT CTGTGTTTCTGTTCCCCCCCAAACCTAAGGATACCCTGATGATCTCTCGGACCCCT GAGGTGACCTGCGTGGTGGTGGACGTGTCACAAGAGGATCCTGAAGTGCAGTTTA ATTGGTACGTGGACGGCGTGGAAGTGCATAACGCCAAAACCAAACCACGCGAAG AACAGTTCAACTCCACCTACAGAGTCGTCAGCGTGCTGACCGTGCTGCACCAGGA TTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTGTCCAACAAGGGACTGCCTTC TTCCATCGAGAAGACCATCTCCAAGGCCAAGGGCCAGCCTCGGGAGCCTCAGGT GTACACCCTCCCTCCATGCCAGGAAGAGATGACCAAGAACCAGGTGTCCCTGTGG TGTCTGGTCAAGGGGTTCTACCCCTCTGACATCGCCGTGGAGTGGGAGTCTAATG GCCAACCCGAGAACAACTACAAGACCACCCCTCCAGTGCTGGACTCTGATGGATC TTTCTTCCTGTACTCCCGGCTGACAGTGGACAAGAGCAGATGGCAGGAAGGCAAC GTGTTCTCCTGCTCCGTGATGCACGAGGCCCTGCATAACCACTATACCCAGAAGTC TCTGTCTCTGTCTCTGGGATCAGGTGGAGGCGGTAGTGGCGGAGGCGGTTCAGGC GGAGGCGGATCTGCACCTACTTCAAGTTCTACAAAGAAAACACAGCTACAACTGG AGCATTTACTGCTGGATTTACAGATGATTTTGAATGGAATTAATAATTACAAGAATC CCAAACTCACCAGGATGCTCACATTTAAGTTTTACATGCCCAAGAAGGCCACAGA ACTGAAACATCTTCAGTGTCTAGAAGAAGAACTCAAACCTCTGGAGGAAGTGCTA AATTTAGCTCAAAGCAAAAACTTTCACTTAAGACCCAGGGACTTAATCAGCAATAT CAACGTAATAGTTCTGGAACTAAAGGGATCTGAAACAACATTCATGTGTGAATATG CTGATGAGACAGCAACCATTGTAGAATTTCTGAACAGATGGATTACCTTTTGTCAA AGCATCATCTCAACACTGACT Amino acid sequence of polypeptide chain H2b (SEQ ID NO: 20) QVQLQQWGAGLLKPSETLSLTCAVYGGSFSDYYWNWIRQPPGKGLEWIGEINHR GSTNSNPSLKSRVTLSLDTSKNQFSLKLRSVTAADTAVYYCAFGYSDYEYNWFDPWG QGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSG VHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCP PCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEV HNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKG QPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVL DSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGSGGGGSGG GGSGGGGSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTFKFYMPKK ATELKHLQCLEEELKPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYA DETATIVEFLNRWITFCQSIISTLT DNA sequence of polypeptide chain H4 (SEQ ID NO: 21) GAGGTGCAGCTGGTGGAAAGCGGAGGAGGCCTGGTACAGCCTGGCGGCTCT CTGCGGCTGTCTTGTGCCGCTTCTGGCTTTACCTTCTCCGATTCCTGGATCCACTGG GTCCGCCAGGCCCCGGGCAAGGGCCTGGAATGGGTGGCTTGGATCTCTCCTTACG GCGGATCTACATATTACGCCGACTCTGTGAAGGGCAGATTCACCATCTCTGCCGAT ACCTCTAAGAACACCGCCTACCTGCAGATGAACAGCCTGAGAGCCGAGGACACC GCTGTGTACTACTGCGCCCGGAGACACTGGCCTGGCGGCTTCGACTACTGGGGCC AAGGCACCCTCGTCACAGTGTCCTCCGCCTCTACCAAGGGACCTAGTGTGTTCCC TCTGGCCCCCTCTTCCAAAAGCACCTCCGGTGGCACCGCTGCTCTGGGCTGCCTG GTGAAGGACTACTTCCCCGAGCCCGTGACCGTGTCTTGGAACTCCGGCGCTCTGA CATCTGGCGTCCATACATTTCCTGCTGTGCTTCAGTCCAGCGGACTGTACAGCCTG TCCTCCGTGGTGACCGTGCCCTCCTCTTCCCTCGGCACACAGACCTATATCTGCAA CGTGAACCACAAGCCTTCCAATACCAAGGTGGACAAGAAAGTGGAACCTAAGTC TTGCGACAAGACCCACACCTGCCCTCCTTGTCCTGCTCCCGAGCTGCTGGGCGGC CCTTCCGTGTTTCTGTTCCCACCTAAGCCCAAGGACACCCTGATGATCAGCCGGAC CCCTGAAGTGACATGCGTGGTCGTGGACGTGTCTCATGAGGATCCTGAAGTGAAG TTCAACTGGTACGTGGATGGCGTGGAAGTGCACAACGCCAAGACCAAACCTCGG GAAGAGCAGTACGCCTCTACCTACAGAGTGGTGTCCGTGCTGACAGTGCTGCACC AGGACTGGCTGAACGGAAAAGAGTACAAGTGCAAGGTGTCCAACAAGGCCCTGC CTGCTCCTATCGAGAAGACCATCTCCAAGGCTAAGGGCCAGCCTAGAGAACCACA AGTGTACACCCTGCCTCCATCTAGAGAGGAGATGACCAAAAACCAGGTTAGTCTG ACCTGTCTGGTCAAGGGCTTCTACCCTTCTGACATCGCCGTGGAGTGGGAGTCCA ACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACTCCGACG GCTCCTTCTTCCTGTACTCCAAGCTGACCGTGGATAAGTCCAGGTGGCAGCAGGG CAACGTGTTCTCCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACCCAG AAGTCCCTGTCTCTGTCCCCTGGCAAG Amino acid sequence of polypeptide chain H4 (SEQ ID NO: 22) EVQLVESGGGLVQPGGSLRLSCAASGFTFSDSWIHWVRQAPGKGLEWVAWISPY GGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARRHWPGGFDYWG QGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSG VHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTH TCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV EVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKA KGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPP VLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK DNA sequence of polypeptide chain L2 (SEQ ID NO: 23) GACATCCAGATGACCCAGTCTCCATCTTCCCTCTCTGCTTCTGTGGGCGACAG AGTGACAATCACCTGCAGAGCCTCTCAGGACGTGTCCACAGCTGTGGCTTGGTAC CAGCAGAAGCCTGGCAAGGCTCCCAAGCTGCTGATCTACTCCGCCAGCTTCCTGT ACTCCGGCGTGCCCTCCCGCTTTTCTGGCTCTGGCTCCGGCACCGACTTCACCCTG ACCATCTCCAGCCTGCAGCCTGAAGATTTTGCTACCTACTACTGCCAGCAGTACCT GTACCACCCTGCCACCTTCGGCCAAGGCACCAAGGTTGAAATCAAGCGGACCGTG GCCGCTCCCAGCGTGTTCATCTTCCCTCCTTCTGATGAACAGCTGAAGTCCGGAAC CGCCTCCGTGGTGTGCCTGCTGAACAACTTCTACCCTCGGGAGGCCAAAGTCCAG TGGAAGGTGGACAACGCCCTGCAGTCCGGAAATAGCCAAGAGTCCGTCACCGAG CAGGATTCTAAGGACAGCACCTATTCTCTGTCCTCCACCCTGACACTGTCCAAGGC CGACTACGAGAAGCACAAGGTGTACGCCTGCGAGGTGACCCATCAGGGCCTGTCT TCTCCTGTGACCAAATCCTTCAACAGAGGCGAGTGT Amino acid sequence of polypeptide chain L2 (SEQ ID NO: 24) DIQMTQSPSSLSASVGDRVTITCRASQDVSTAVAWYQQKPGKAPKLLIYSASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYLYHPATFGQGTKVEIKRTVAAPSV FIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDST YSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC DNA sequence of polypeptide chain H5 (SEQ ID NO: 25) GAGGTGCAGCTGGTGGAAAGCGGAGGAGGCCTGGTACAGCCTGGCGGCTCT CTGCGGCTGTCTTGTGCCGCTTCTGGCTTTACCTTCTCCGATTCCTGGATCCACTGG GTCCGCCAGGCCCCGGGCAAGGGCCTGGAATGGGTGGCTTGGATCTCTCCTTACG GCGGATCTACATATTACGCCGACTCTGTGAAGGGCAGATTCACCATCTCTGCCGAT ACCTCTAAGAACACCGCCTACCTGCAGATGAACAGCCTGAGAGCCGAGGACACC GCTGTGTACTACTGCGCCCGGAGACACTGGCCTGGCGGCTTCGACTACTGGGGCC AAGGCACCCTCGTCACAGTGTCCTCCGCCTCTACCAAGGGACCTAGTGTGTTCCC TCTGGCCCCCTCTTCCAAAAGCACCTCCGGTGGCACCGCTGCTCTGGGCTGCCTG GTGAAGGACTACTTCCCCGAGCCCGTGACCGTGTCTTGGAACTCCGGCGCTCTGA CATCTGGCGTCCATACATTTCCTGCTGTGCTTCAGTCCAGCGGACTGTACAGCCTG TCCTCCGTGGTGACCGTGCCCTCCTCTTCCCTCGGCACACAGACCTATATCTGCAA CGTGAACCACAAGCCTTCCAATACCAAGGTGGACAAGAAAGTGGAACCTAAGTC TTGCGACAAGACCCACACCTGCCCTCCTTGTCCTGCTCCCGAGCTGCTGGGCGGC CCTTCCGTGTTTCTGTTCCCACCTAAGCCCAAGGACACCCTGATGATCAGCCGGAC CCCTGAAGTGACATGCGTGGTCGTGGACGTGTCTCATGAGGATCCTGAAGTGAAG TTCAACTGGTACGTGGATGGCGTGGAAGTGCACAACGCCAAGACCAAACCTCGG GAAGAGCAGTACGCCTCTACCTACAGAGTGGTGTCCGTGCTGACAGTGCTGCACC AGGACTGGCTGAACGGAAAAGAGTACAAGTGCAAGGTGTCCAACAAGGCCCTGC CTGCTCCTATCGAGAAGACCATCTCCAAGGCTAAGGGCCAGCCTAGAGAACCACA AGTGTACACCCTGCCTCCATGTAGAGAGGAGATGACCAAAAACCAGGTTAGTCTG TGGTGTCTGGTCAAGGGCTTCTACCCTTCTGACATCGCCGTGGAGTGGGAGTCCA ACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACTCCGACG GCTCCTTCTTCCTGTACTCCAAGCTGACCGTGGATAAGTCCAGGTGGCAGCAGGG CAACGTGTTCTCCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACCCAG AAGTCCCTGTCTCTGTCCCCTGGC Amino acid sequence of polypeptide chain H5 (SEQ ID NO: 26) EVQLVESGGGLVQPGGSLRLSCAASGFTFSDSWIHWVRQAPGKGLEWVAWISPY GGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARRHWPGGFDYWG QGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSG VHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTH TCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV EVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKA KGQPREPQVYTLPPCREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPP VLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG DNA sequence of polypeptide chain H6 (SEQ ID NO: 27) GAGGTGCAGCTGGTGGAAAGCGGAGGAGGCCTGGTACAGCCTGGCGGCTCT CTGCGGCTGTCTTGTGCCGCTTCTGGCTTTACCTTCTCCGATTCCTGGATCCACTGG GTCCGCCAGGCCCCGGGCAAGGGCCTGGAATGGGTGGCTTGGATCTCTCCTTACG GCGGATCTACATATTACGCCGACTCTGTGAAGGGCAGATTCACCATCTCTGCCGAT ACCTCTAAGAACACCGCCTACCTGCAGATGAACAGCCTGAGAGCCGAGGACACC GCTGTGTACTACTGCGCCCGGAGACACTGGCCTGGCGGCTTCGACTACTGGGGCC AAGGCACCCTCGTCACAGTGTCCTCCGCCTCTACCAAGGGACCTAGTGTGTTCCC TCTGGCCCCCTCTTCCAAAAGCACCTCCGGTGGCACCGCTGCTCTGGGCTGCCTG GTGAAGGACTACTTCCCCGAGCCCGTGACCGTGTCTTGGAACTCCGGCGCTCTGA CATCTGGCGTCCATACATTTCCTGCTGTGCTTCAGTCCAGCGGACTGTACAGCCTG TCCTCCGTGGTGACCGTGCCCTCCTCTTCCCTCGGCACACAGACCTATATCTGCAA CGTGAACCACAAGCCTTCCAATACCAAGGTGGACAAGAAAGTGGAACCTAAGTC TTGCGACAAGACCCACACCTGCCCTCCTTGTCCTGCTCCCGAGCTGCTGGGCGGC CCTTCCGTGTTTCTGTTCCCACCTAAGCCCAAGGACACCCTGATGATCAGCCGGAC CCCTGAAGTGACATGCGTGGTCGTGGACGTGTCTCATGAGGATCCTGAAGTGAAG TTCAACTGGTACGTGGATGGCGTGGAAGTGCACAACGCCAAGACCAAACCTCGG GAAGAGCAGTACGCCTCTACCTACAGAGTGGTGTCCGTGCTGACAGTGCTGCACC AGGACTGGCTGAACGGAAAAGAGTACAAGTGCAAGGTGTCCAACAAGGCCCTGC CTGCTCCTATCGAGAAGACCATCTCCAAGGCTAAGGGCCAGCCTAGAGAACCACA AGTGTGCACCCTGCCTCCATCTAGAGAGGAGATGACCAAAAACCAGGTTAGTCTG AGCTGTGCTGTCAAGGGCTTCTACCCTTCTGACATCGCCGTGGAGTGGGAGTCCA ACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACTCCGACG GCTCCTTCTTCCTGGTCTCCAAGCTGACCGTGGATAAGTCCAGGTGGCAGCAGGG CAACGTGTTCTCCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACCCAG AAGTCCCTGTCTCTGTCCCCTGGC Amino acid sequence of polypeptide chain H6 (SEQ ID NO: 28) EVQLVESGGGLVQPGGSLRLSCAASGFTFSDSWIHWVRQAPGKGLEWVAWISPY GGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARRHWPGGFDYWG QGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSG VHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTH TCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV EVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKA KGQPREPQVCTLPPSREEMTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPP VLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG DNA sequence of polypeptide chain H5a (SEQ ID NO: 29) GAGGTGCAGCTGGTGGAAAGCGGAGGAGGCCTGGTACAGCCTGGCGGCTCT CTGCGGCTGTCTTGTGCCGCTTCTGGCTTTACCTTCTCCGATTCCTGGATCCACTGG GTCCGCCAGGCCCCGGGCAAGGGCCTGGAATGGGTGGCTTGGATCTCTCCTTACG GCGGATCTACATATTACGCCGACTCTGTGAAGGGCAGATTCACCATCTCTGCCGAT ACCTCTAAGAACACCGCCTACCTGCAGATGAACAGCCTGAGAGCCGAGGACACC GCTGTGTACTACTGCGCCCGGAGACACTGGCCTGGCGGCTTCGACTACTGGGGCC AAGGCACCCTCGTCACAGTGTCCTCCGCCTCTACCAAGGGACCTAGTGTGTTCCC TCTGGCCCCCTCTTCCAAAAGCACCTCCGGTGGCACCGCTGCTCTGGGCTGCCTG GTGAAGGACTACTTCCCCGAGCCCGTGACCGTGTCTTGGAACTCCGGCGCTCTGA CATCTGGCGTCCATACATTTCCTGCTGTGCTTCAGTCCAGCGGACTGTACAGCCTG TCCTCCGTGGTGACCGTGCCCTCCTCTTCCCTCGGCACACAGACCTATATCTGCAA CGTGAACCACAAGCCTTCCAATACCAAGGTGGACAAGAAAGTGGAACCTAAGTC TTGCGACAAGACCCACACCTGCCCTCCTTGTCCTGCTCCCGAGCTGCTGGGCGGC CCTTCCGTGTTTCTGTTCCCACCTAAGCCCAAGGACACCCTGATGATCAGCCGGAC CCCTGAAGTGACATGCGTGGTCGTGGACGTGTCTCATGAGGATCCTGAAGTGAAG TTCAACTGGTACGTGGATGGCGTGGAAGTGCACAACGCCAAGACCAAACCTCGG GAAGAGCAGTACGCCTCTACCTACAGAGTGGTGTCCGTGCTGACAGTGCTGCACC AGGACTGGCTGAACGGAAAAGAGTACAAGTGCAAGGTGTCCAACAAGGCCCTGC CTGCTCCTATCGAGAAGACCATCTCCAAGGCTAAGGGCCAGCCTAGAGAACCACA AGTGTACACCCTGCCTCCATGTAGAGAGGAGATGACCAAAAACCAGGTTAGTCTG TGGTGTCTGGTCAAGGGCTTCTACCCTTCTGACATCGCCGTGGAGTGGGAGTCCA ACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACTCCGACG GCTCCTTCTTCCTGTACTCCAAGCTGACCGTGGATAAGTCCAGGTGGCAGCAGGG CAACGTGTTCTCCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACCCAG AAGTCCCTGTCTCTGTCCCCTGGCTCAGGTGGAGGCGGTAGTGGCGGAGGCGGTT CAGGCGGAGGCGGATCTATTTGGGAGCTGAAGAAAGACGTGTACGTGGTCGAGC TGGACTGGTACCCTGATGCCCCAGGCGAGATGGTCGTGCTGACCTGCGATACACC AGAGGAAGATGGTATCACCTGGACACTGGATCAGTCCTCAGAGGTGCTGGGCTCT GGTAAAACACTGACCATTCAGGTGAAGGAGTTCGGTGACGCTGGACAGTACACTT GTCATAAGGGCGGGGAGGTGCTGTCTCACTCCCTGCTGCTGCTGCATAAGAAGGA GGATGGAATCTGGTCCACTGACATCCTGAAAGACCAGAAGGAGCCAAAGAACAA AACCTTCCTGCGATGCGAGGCTAAGAACTACAGCGGCCGCTTTACATGCTGGTGG CTGACAACCATCAGCACCGATCTGACCTTTAGCGTGAAGTCATCCAGGGGCAGTT CAGACCCTCAGGGAGTCACATGTGGCGCCGCAACCCTGTCAGCAGAGCGAGTGC GGGGAGACAATAAGGAATACGAGTACAGCGTCGAGTGTCAGGAGGATTCCGCATG TCCAGCTGCAGAAGAATCCCTGCCTATCGAAGTCATGGTGGACGCTGTGCATAAA CTGAAGTACGAGAATTACACCAGCAGCTTTTTCATCCGGGACATCATCAAGCCCGA TCCACCTAAGAATCTGCAGCTGAAGCCTCTGAAAAATAGCCGACAGGTCGAAGTG TCATGGGAATACCCAGACACCTGGTCAACACCACACTCCTACTTCTCCCTGACCTT CTGTGTGCAGGTCCAGGGAAAAAGCAAGCGGGAAAAGAAAGATCGGGTGTTCAC CGACAAGACCAGTGCTACAGTGATTTGCCGGAAGAATGCCAGCATTTCTGTCAGA GCTCAGGACCGGTACTATAGCTCTTCCTGGAGCGAGTGGGCTTCAGTGCCATGTTC TGGAGGCGGTGGATCTGGCGGAGGTGGAAGCGGAGGCGGTGGATCTAGAAACCT GCCCGTCGCAACCCCTGATCCAGGGATGTTCCCCTGTCTGCATCACAGCCAGAATC TGCTGAGGGCTGTCTCCAACATGCTGCAGAAGGCTCGACAGACCCTGGAGTTCTA CCCATGTACCAGCGAAGAGATCGACCACGAGGATATCACAAAGGATAAAACCAGC ACAGTGGAAGCATGCCTGCCTCTGGAACTGACCAAGAATGAGAGCTGCCTGAATA GCAGGGAGACCTCCTTCATCACCAACGGCTCATGCCTGGCTTCAAGGAAGACCAG CTTCATGATGGCTCTGTGTCTGAGCTCTATCTATGAGGACCTGAAGATGTACCAGG TGGAGTTCAAGACCATGAACGCCAAGCTGCTGATGGATCCAAAGAGGCAGATCTT CCTGGATCAGAATATGCTGGCAGTGATCGATGAGCTGATGCAGGCCCTGAATTTTA ACAGTGAGACAGTGCCTCAGAAGAGCTCTCTGGAAGAGCCAGACTTTTACAAAA CTAAGATCAAGCTGTGCATTCTGCTGCACGCTTTCCGCATCAGAGCTGTCACTATC GATAGAGTGATGAGCTATCTGAATGCCTCA Amino acid sequence of polypeptide chain H5a (SEQ ID NO: 30) EVQLVESGGGLVQPGGSLRLSCAASGFTFSDSWIHWVRQAPGKGLEWVAWISPY GGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARRHWPGGFDYWG QGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSG VHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTH TCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV EVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKA KGQPREPQVYTLPPCREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPP VLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGSGGGGS GGGGSGGGGSIWELKKDVYVVELDWYPDAPGEMVVLTCDTPEEDGITWTLDQSSE VLGSGKTLTIQVKEFGDAGQYTCHKGGEVLSHSLLLLHKKEDGIWSTDILKDQKEPK NKTFLRCEAKNYSGRFTCWWLTTISTDLTFSVKSSRGSSDPQGVTCGAATLSAERVRG DNKEYEYSVECQEDSACPAAEESLPIEVMVDAVHKLKYENYTSSFFIRDIIKPDPPKNL QLKPLKNSRQVEVSWEYPDTWSTPHSYFSLTFCVQVQGKSKREKKDRVFTDKTSATV ICRKNASISVRAQDRYYSSSWSEWASVPCSGGGGSGGGGSGGGGSRNLPVATPDPGM FPCLHHSQNLLRAVSNMLQKARQTLEFYPCTSEEIDHEDITKDKTSTVEACLPLELTK NESCLNSRETSFITNGSCLASRKTSFMMALCLSSIYEDLKMYQVEFKTMNAKLLMDP KRQIFLDQNMLAVIDELMQALNFNSETVPQKSSLEEPDFYKTKIKLCILLHAFRIRAVT IDRVMSYLNAS DNA sequence of polypeptide chain H6a (SEQ ID NO: 31) GAGGTGCAGCTGGTGGAAAGCGGAGGAGGCCTGGTACAGCCTGGCGGCTCT CTGCGGCTGTCTTGTGCCGCTTCTGGCTTTACCTTCTCCGATTCCTGGATCCACTGG GTCCGCCAGGCCCCGGGCAAGGGCCTGGAATGGGTGGCTTGGATCTCTCCTTACG GCGGATCTACATATTACGCCGACTCTGTGAAGGGCAGATTCACCATCTCTGCCGAT ACCTCTAAGAACACCGCCTACCTGCAGATGAACAGCCTGAGAGCCGAGGACACC GCTGTGTACTACTGCGCCCGGAGACACTGGCCTGGCGGCTTCGACTACTGGGGCC AAGGCACCCTCGTCACAGTGTCCTCCGCCTCTACCAAGGGACCTAGTGTGTTCCC TCTGGCCCCCTCTTCCAAAAGCACCTCCGGTGGCACCGCTGCTCTGGGCTGCCTG GTGAAGGACTACTTCCCCGAGCCCGTGACCGTGTCTTGGAACTCCGGCGCTCTGA CATCTGGCGTCCATACATTTCCTGCTGTGCTTCAGTCCAGCGGACTGTACAGCCTG TCCTCCGTGGTGACCGTGCCCTCCTCTTCCCTCGGCACACAGACCTATATCTGCAA CGTGAACCACAAGCCTTCCAATACCAAGGTGGACAAGAAAGTGGAACCTAAGTC TTGCGACAAGACCCACACCTGCCCTCCTTGTCCTGCTCCCGAGCTGCTGGGCGGC CCTTCCGTGTTTCTGTTCCCACCTAAGCCCAAGGACACCCTGATGATCAGCCGGAC CCCTGAAGTGACATGCGTGGTCGTGGACGTGTCTCATGAGGATCCTGAAGTGAAG TTCAACTGGTACGTGGATGGCGTGGAAGTGCACAACGCCAAGACCAAACCTCGG GAAGAGCAGTACGCCTCTACCTACAGAGTGGTGTCCGTGCTGACAGTGCTGCACC AGGACTGGCTGAACGGAAAAGAGTACAAGTGCAAGGTGTCCAACAAGGCCCTGC CTGCTCCTATCGAGAAGACCATCTCCAAGGCTAAGGGCCAGCCTAGAGAACCACA AGTGTGCACCCTGCCTCCATCTAGAGAGGAGATGACCAAAAACCAGGTTAGTCTG AGCTGTGCTGTCAAGGGCTTCTACCCTTCTGACATCGCCGTGGAGTGGGAGTCCA ACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACTCCGACG GCTCCTTCTTCCTGGTCTCCAAGCTGACCGTGGATAAGTCCAGGTGGCAGCAGGG CAACGTGTTCTCCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACCCAG AAGTCCCTGTCTCTGTCCCCTGGCTCAGGTGGAGGCGGTAGTGGCGGAGGCGGTT CAGGCGGAGGCGGATCTGCACCTACTTCAAGTTCTACAAAGAAAACACAGCTACA ACTGGAGCATTTACTGCTGGATTTACAGATGATTTTGAATGGAATTAATAATTACAA GAATCCCAAACTCACCAGGATGCTCACATTTAAGTTTTACATGCCCAAGAAGGCC ACAGAACTGAAACATCTTCAGTGTCTAGAAGAAGAACTCAAACCTCTGGAGGAA GTGCTAAATTTAGCTCAAAGCAAAAACTTTCACTTAAGACCCAGGGACTTAATCA GCAATATCAACGTAATAGTTCTGGAACTAAAGGGATCTGAAACAACATTCATGTGT GAATATGCTGATGAGACAGCAACCATTGTAGAATTTCTGAACAGATGGATTACCTT TTGTCAAAGCATCATCTCAACACTGACT Amino acid sequence of polypeptide chain H6a (SEQ ID NO: 32) EVQLVESGGGLVQPGGSLRLSCAASGFTFSDSWIHWVRQAPGKGLEWVAWISPY GGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARRHWPGGFDYWG QGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSG VHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTH TCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV EVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKA KGQPREPQVCTLPPSREEMTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPP VLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGSGGGGS GGGGSGGGGSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTFKFYMP KKATELKHLQCLEEELKPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCE YADETATIVEFLNRWITFCQSIISTLT DNA sequence of polypeptide chain H5b (SEQ ID NO: 33) GAGGTGCAGCTGGTGGAAAGCGGAGGAGGCCTGGTACAGCCTGGCGGCTCT CTGCGGCTGTCTTGTGCCGCTTCTGGCTTTACCTTCTCCGATTCCTGGATCCACTGG GTCCGCCAGGCCCCGGGCAAGGGCCTGGAATGGGTGGCTTGGATCTCTCCTTACG GCGGATCTACATATTACGCCGACTCTGTGAAGGGCAGATTCACCATCTCTGCCGAT ACCTCTAAGAACACCGCCTACCTGCAGATGAACAGCCTGAGAGCCGAGGACACC GCTGTGTACTACTGCGCCCGGAGACACTGGCCTGGCGGCTTCGACTACTGGGGCC AAGGCACCCTCGTCACAGTGTCCTCCGCCTCTACCAAGGGACCTAGTGTGTTCCC TCTGGCCCCCTCTTCCAAAAGCACCTCCGGTGGCACCGCTGCTCTGGGCTGCCTG GTGAAGGACTACTTCCCCGAGCCCGTGACCGTGTCTTGGAACTCCGGCGCTCTGA CATCTGGCGTCCATACATTTCCTGCTGTGCTTCAGTCCAGCGGACTGTACAGCCTG TCCTCCGTGGTGACCGTGCCCTCCTCTTCCCTCGGCACACAGACCTATATCTGCAA CGTGAACCACAAGCCTTCCAATACCAAGGTGGACAAGAAAGTGGAACCTAAGTC TTGCGACAAGACCCACACCTGCCCTCCTTGTCCTGCTCCCGAGCTGCTGGGCGGC CCTTCCGTGTTTCTGTTCCCACCTAAGCCCAAGGACACCCTGATGATCAGCCGGAC CCCTGAAGTGACATGCGTGGTCGTGGACGTGTCTCATGAGGATCCTGAAGTGAAG TTCAACTGGTACGTGGATGGCGTGGAAGTGCACAACGCCAAGACCAAACCTCGG GAAGAGCAGTACGCCTCTACCTACAGAGTGGTGTCCGTGCTGACAGTGCTGCACC AGGACTGGCTGAACGGAAAAGAGTACAAGTGCAAGGTGTCCAACAAGGCCCTGC CTGCTCCTATCGAGAAGACCATCTCCAAGGCTAAGGGCCAGCCTAGAGAACCACA AGTGTACACCCTGCCTCCATGTAGAGAGGAGATGACCAAAAACCAGGTTAGTCTG TGGTGTCTGGTCAAGGGCTTCTACCCTTCTGACATCGCCGTGGAGTGGGAGTCCA ACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACTCCGACG GCTCCTTCTTCCTGTACTCCAAGCTGACCGTGGATAAGTCCAGGTGGCAGCAGGG CAACGTGTTCTCCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACCCAG AAGTCCCTGTCTCTGTCCCCTGGCTCAGGTGGAGGCGGTAGTGGCGGAGGCGGTT CAGGCGGAGGCGGATCTGCACCTACTTCAAGTTCTACAAAGAAAACACAGCTACA ACTGGAGCATTTACTGCTGGATTTACAGATGATTTTGAATGGAATTAATAATTACAA GAATCCCAAACTCACCAGGATGCTCACATTTAAGTTTTACATGCCCAAGAAGGCC ACAGAACTGAAACATCTTCAGTGTCTAGAAGAAGAACTCAAACCTCTGGAGGAA GTGCTAAATTTAGCTCAAAGCAAAAACTTTCACTTAAGACCCAGGGACTTAATCA GCAATATCAACGTAATAGTTCTGGAACTAAAGGGATCTGAAACAACATTCATGTGT GAATATGCTGATGAGACAGCAACCATTGTAGAATTTCTGAACAGATGGATTACCTT TTGTCAAAGCATCATCTCAACACTGACT Amino acid sequence of polypeptide chain H5b (SEQ ID NO: 34) EVQLVESGGGLVQPGGSLRLSCAASGFTFSDSWIHWVRQAPGKGLEWVAWISPY GGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARRHWPGGFDYWG QGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSG VHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTH TCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV EVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKA KGQPREPQVYTLPPCREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPP VLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGSGGGGS GGGGSGGGGSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTFKFYMP KKATELKHLQCLEEELKPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCE YADETATIVEFLNRWITFCQSIISTLT DNA sequence of polypeptide chain H7 (SEQ ID NO: 35) GAAGTGCAGCTGCAGCAGTCCGGCCCCGGACTGGTCAAACCCTCCCAGACCC TGTCTCTTACCTGCGCCATCTCCGGCGACTCTGTGTCCAGCAACTCTGCCGCTTGG AATTGGATCCGGCAGTCTCCTAGTAGAGGCCTGGAATGGCTGGGCAAGACATACT ACCGGTTCAAGTGGTATTCCGATTACGCCGTGTCCGTGAAGGGCAGAATCACCATC AACCCTGATACCTCTAAGAATCAATTCTCCCTGCAGCTGAACTCCGTGACCCCTGA GGACACCGCTGTGTTTTACTGCACCAGAGAGAGCACCACCTACGACCTGCTGGCT GGCCCTTTCGACTACTGGGGCCAGGGAACACTCGTCACCGTGTCTTCCGCCTCTA CAAAGGGCCCATCCGTTTTTCCTCTGGCCCCGTCCTCTAAGTCTACCTCTGGCGGC ACCGCCGCTCTGGGCTGCCTGGTGAAGGACTACTTCCCCGAGCCTGTGACCGTGT CCTGGAACAGCGGCGCTCTGACCTCCGGCGTGCATACCTTCCCTGCTGTTCTGCAA TCTTCTGGCCTGTACTCTCTGTCCTCTGTGGTGACCGTGCCTTCCAGCTCTCTGGG CACCCAGACCTACATCTGCAACGTGAACCATAAGCCTTCTAACACTAAGGTGGAC AAGAAAGTGGAACCCAAGTCCTGCGACAAAACCCACACCTGTCCTCCTTGTCCTG CCCCTGAGCTGCTGGGCGGACCTAGCGTGTTCCTGTTCCCACCTAAGCCTAAGGAT ACCCTGATGATCTCCCGGACCCCTGAAGTGACCTGCGTGGTCGTGGACGTGTCTC ACGAAGATCCTGAAGTCAAGTTCAACTGGTACGTGGATGGCGTCGAGGTGCACAA CGCCAAGACCAAGCCCCGGGAAGAGCAGTACAACTCCACATATAGAGTGGTGAG TGTGCTGACAGTACTGCACCAGGACTGGCTGAACGGCAAAGAGTACAAGTGCAA GGTGTCCAACAAGGCTCTGCCTGCTCCTATCGAGAAGACAATCTCCAAGGCCAAG GGACAGCCTAGAGAACCTCAAGTGTACACCCTCCCACCTTCTCGCGAGGAGATGA CCAAGAACCAGGTGAGCCTGACCTGTCTGGTGAAAGGCTTCTACCCCTCCGACAT CGCCGTGGAGTGGGAGTCCAATGGCCAGCCAGAGAACAACTACAAGACCACACC TCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTGTACTCCAAGCTGACCGTGGAC AAGTCTAGATGGCAGCAGGGCAACGTGTTTAGCTGCTCCGTGATGCACGAGGCCC TGCACAACCACTACACCCAGAAGTCTCTGTCCCTGAGCCCTGGAAAA Amino acid sequence of polypeptide chain H7 (SEQ ID NO: 36) EVQLQQSGPGLVKPSQTLSLTCAISGDSVSSNSAAWNWIRQSPSRGLEWLGKTYY RFKWYSDYAVSVKGRITINPDTSKNQFSLQLNSVTPEDTAVFYCTRESTTYDLLAGPF DYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKS CDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNW YVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIE KTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNY KTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK DNA sequence of polypeptide chain L3 (SEQ ID NO: 37) GATATCGTGATGACCCAGTCCCCCGATTCTCTGGCCGTGTCCCTGGGCGAGCG GGCTACAATCAACTGCAAGTCCTCTCAGACCGTGCTGTACTCTTCCAACAACAAG AAGTACCTGGCTTGGTACCAGCAGAAGCCTGGCCAGCCTCCTAATCTGCTGATCTA CTGGGCCTCTACAAGGGAGTCCGGCGTCCCCGATAGATTTAGCGGATCCGGCTCTG GAACCGACTTCACCCTGACCATCTCCAGCCTGCAAGCCGAGGACGTGGCTGTGTA CTACTGCCAGCAGTACTATTCTACTCCTTTTACCTTCGGCCCCGGCACCAAAGTTG AGATCAAGCGGACCGTGGCCGCTCCTTCCGTGTTCATCTTCCCTCCATCCGACGAA CAGCTGAAGTCCGGCACCGCTAGCGTGGTGTGCCTGCTGAACAACTTCTACCCTA GAGAAGCCAAAGTGCAGTGGAAGGTGGACAACGCCCTGCAGTCTGGCAACTCCC AAGAGTCTGTCACCGAGCAGGACTCTAAGGACTCCACCTACTCCCTCAGCTCCAC ACTGACCCTGTCCAAGGCCGACTACGAGAAGCACAAGGTGTACGCCTGCGAAGT GACCCACCAGGGCCTGTCTTCTCCTGTGACCAAGTCCTTCAACAGAGGCGAATGT Amino acid sequence of polypeptide chain L3 (SEQ ID NO: 38) DIVMTQSPDSLAVSLGERATINCKSSQTVLYSSNNKKYLAWYQQKPGQPPNLLIY WASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQYYSTPFTFGPGTKVEIKR TVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTE QDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC DNA sequence of polypeptide chain H8 (SEQ ID NO: 39) GAAGTGCAGCTGCAGCAGTCCGGCCCCGGACTGGTCAAACCCTCCCAGACCC TGTCTCTTACCTGCGCCATCTCCGGCGACTCTGTGTCCAGCAACTCTGCCGCTTGG AATTGGATCCGGCAGTCTCCTAGTAGAGGCCTGGAATGGCTGGGCAAGACATACT ACCGGTTCAAGTGGTATTCCGATTACGCCGTGTCCGTGAAGGGCAGAATCACCATC AACCCTGATACCTCTAAGAATCAATTCTCCCTGCAGCTGAACTCCGTGACCCCTGA GGACACCGCTGTGTTTTACTGCACCAGAGAGAGCACCACCTACGACCTGCTGGCT GGCCCTTTCGACTACTGGGGCCAGGGAACACTCGTCACCGTGTCTTCCGCCTCTA CAAAGGGCCCATCCGTTTTTCCTCTGGCCCCGTCCTCTAAGTCTACCTCTGGCGGC ACCGCCGCTCTGGGCTGCCTGGTGAAGGACTACTTCCCCGAGCCTGTGACCGTGT CCTGGAACAGCGGCGCTCTGACCTCCGGCGTGCATACCTTCCCTGCTGTTCTGCAA TCTTCTGGCCTGTACTCTCTGTCCTCTGTGGTGACCGTGCCTTCCAGCTCTCTGGG CACCCAGACCTACATCTGCAACGTGAACCATAAGCCTTCTAACACTAAGGTGGAC AAGAAAGTGGAACCCAAGTCCTGCGACAAAACCCACACCTGTCCTCCTTGTCCTG CCCCTGAGCTGCTGGGCGGACCTAGCGTGTTCCTGTTCCCACCTAAGCCTAAGGAT ACCCTGATGATCTCCCGGACCCCTGAAGTGACCTGCGTGGTCGTGGACGTGTCTC ACGAAGATCCTGAAGTCAAGTTCAACTGGTACGTGGATGGCGTCGAGGTGCACAA CGCCAAGACCAAGCCCCGGGAAGAGCAGTACAACTCCACATATAGAGTGGTGAG TGTGCTGACAGTACTGCACCAGGACTGGCTGAACGGCAAAGAGTACAAGTGCAA GGTGTCCAACAAGGCTCTGCCTGCTCCTATCGAGAAGACAATCTCCAAGGCCAAG GGACAGCCTAGAGAACCTCAAGTGTACACCCTCCCACCTTGTCGCGAGGAGATGA CCAAGAACCAGGTGAGCCTGTGGTGTCTGGTGAAAGGCTTCTACCCCTCCGACAT CGCCGTGGAGTGGGAGTCCAATGGCCAGCCAGAGAACAACTACAAGACCACACC TCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTGTACTCCAAGCTGACCGTGGAC AAGTCTAGATGGCAGCAGGGCAACGTGTTTAGCTGCTCCGTGATGCACGAGGCCC TGCACAACCACTACACCCAGAAGTCTCTGTCCCTGAGCCCTGGA Amino acid sequence of polypeptide chain H8 (SEQ ID NO: 40) EVQLQQSGPGLVKPSQTLSLTCAISGDSVSSNSAAWNWIRQSPSRGLEWLGKTYY RFKWYSDYAVSVKGRITINPDTSKNQFSLQLNSVTPEDTAVFYCTRESTTYDLLAGPF DYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKS CDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNW YVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIE KTISKAKGQPREPQVYTLPPCREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG DNA sequence of polypeptide chain H9 (SEQ ID NO: 41) GAAGTGCAGCTGCAGCAGTCCGGCCCCGGACTGGTCAAACCCTCCCAGACCC TGTCTCTTACCTGCGCCATCTCCGGCGACTCTGTGTCCAGCAACTCTGCCGCTTGG AATTGGATCCGGCAGTCTCCTAGTAGAGGCCTGGAATGGCTGGGCAAGACATACT ACCGGTTCAAGTGGTATTCCGATTACGCCGTGTCCGTGAAGGGCAGAATCACCATC AACCCTGATACCTCTAAGAATCAATTCTCCCTGCAGCTGAACTCCGTGACCCCTGA GGACACCGCTGTGTTTTACTGCACCAGAGAGAGCACCACCTACGACCTGCTGGCT GGCCCTTTCGACTACTGGGGCCAGGGAACACTCGTCACCGTGTCTTCCGCCTCTA CAAAGGGCCCATCCGTTTTTCCTCTGGCCCCGTCCTCTAAGTCTACCTCTGGCGGC ACCGCCGCTCTGGGCTGCCTGGTGAAGGACTACTTCCCCGAGCCTGTGACCGTGT CCTGGAACAGCGGCGCTCTGACCTCCGGCGTGCATACCTTCCCTGCTGTTCTGCAA TCTTCTGGCCTGTACTCTCTGTCCTCTGTGGTGACCGTGCCTTCCAGCTCTCTGGG CACCCAGACCTACATCTGCAACGTGAACCATAAGCCTTCTAACACTAAGGTGGAC AAGAAAGTGGAACCCAAGTCCTGCGACAAAACCCACACCTGTCCTCCTTGTCCTG CCCCTGAGCTGCTGGGCGGACCTAGCGTGTTCCTGTTCCCACCTAAGCCTAAGGAT ACCCTGATGATCTCCCGGACCCCTGAAGTGACCTGCGTGGTCGTGGACGTGTCTC ACGAAGATCCTGAAGTCAAGTTCAACTGGTACGTGGATGGCGTCGAGGTGCACAA CGCCAAGACCAAGCCCCGGGAAGAGCAGTACAACTCCACATATAGAGTGGTGAG TGTGCTGACAGTACTGCACCAGGACTGGCTGAACGGCAAAGAGTACAAGTGCAA GGTGTCCAACAAGGCTCTGCCTGCTCCTATCGAGAAGACAATCTCCAAGGCCAAG GGACAGCCTAGAGAACCTCAAGTGTGCACCCTCCCACCTTCTCGCGAGGAGATGA CCAAGAACCAGGTGAGCCTGTCCTGTGCTGTGAAAGGCTTCTACCCCTCCGACAT CGCCGTGGAGTGGGAGTCCAATGGCCAGCCAGAGAACAACTACAAGACCACACC TCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTGGTGTCCAAGCTGACCGTGGAC AAGTCTAGATGGCAGCAGGGCAACGTGTTTAGCTGCTCCGTGATGCACGAGGCCC TGCACAACCACTACACCCAGAAGTCTCTGTCCCTGAGCCCTGGA Amino acid sequence of polypeptide chain H9 (SEQ ID NO: 42) EVQLQQSGPGLVKPSQTLSLTCAISGDSVSSNSAAWNWIRQSPSRGLEWLGKTYY RFKWYSDYAVSVKGRITINPDTSKNQFSLQLNSVTPEDTAVFYCTRESTTYDLLAGPF DYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKS CDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNW YVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIE KTISKAKGQPREPQVCTLPPSREEMTKNQVSLSCAVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG DNA sequence of polypeptide chain H8a (SEQ ID NO: 43) GAAGTGCAGCTGCAGCAGTCCGGCCCCGGACTGGTCAAACCCTCCCAGACCC TGTCTCTTACCTGCGCCATCTCCGGCGACTCTGTGTCCAGCAACTCTGCCGCTTGG AATTGGATCCGGCAGTCTCCTAGTAGAGGCCTGGAATGGCTGGGCAAGACATACT ACCGGTTCAAGTGGTATTCCGATTACGCCGTGTCCGTGAAGGGCAGAATCACCATC AACCCTGATACCTCTAAGAATCAATTCTCCCTGCAGCTGAACTCCGTGACCCCTGA GGACACCGCTGTGTTTTACTGCACCAGAGAGAGCACCACCTACGACCTGCTGGCT GGCCCTTTCGACTACTGGGGCCAGGGAACACTCGTCACCGTGTCTTCCGCCTCTA CAAAGGGCCCATCCGTTTTTCCTCTGGCCCCGTCCTCTAAGTCTACCTCTGGCGGC ACCGCCGCTCTGGGCTGCCTGGTGAAGGACTACTTCCCCGAGCCTGTGACCGTGT CCTGGAACAGCGGCGCTCTGACCTCCGGCGTGCATACCTTCCCTGCTGTTCTGCAA TCTTCTGGCCTGTACTCTCTGTCCTCTGTGGTGACCGTGCCTTCCAGCTCTCTGGG CACCCAGACCTACATCTGCAACGTGAACCATAAGCCTTCTAACACTAAGGTGGAC AAGAAAGTGGAACCCAAGTCCTGCGACAAAACCCACACCTGTCCTCCTTGTCCTG CCCCTGAGCTGCTGGGCGGACCTAGCGTGTTCCTGTTCCCACCTAAGCCTAAGGAT ACCCTGATGATCTCCCGGACCCCTGAAGTGACCTGCGTGGTCGTGGACGTGTCTC ACGAAGATCCTGAAGTCAAGTTCAACTGGTACGTGGATGGCGTCGAGGTGCACAA CGCCAAGACCAAGCCCCGGGAAGAGCAGTACAACTCCACATATAGAGTGGTGAG TGTGCTGACAGTACTGCACCAGGACTGGCTGAACGGCAAAGAGTACAAGTGCAA GGTGTCCAACAAGGCTCTGCCTGCTCCTATCGAGAAGACAATCTCCAAGGCCAAG GGACAGCCTAGAGAACCTCAAGTGTACACCCTCCCACCTTGTCGCGAGGAGATGA CCAAGAACCAGGTGAGCCTGTGGTGTCTGGTGAAAGGCTTCTACCCCTCCGACAT CGCCGTGGAGTGGGAGTCCAATGGCCAGCCAGAGAACAACTACAAGACCACACC TCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTGTACTCCAAGCTGACCGTGGAC AAGTCTAGATGGCAGCAGGGCAACGTGTTTAGCTGCTCCGTGATGCACGAGGCCC TGCACAACCACTACACCCAGAAGTCTCTGTCCCTGAGCCCTGGATCAGGTGGAGG CGGTAGTGGCGGAGGCGGTTCAGGCGGAGGCGGATCTATTTGGGAGCTGAAGAA AGACGTGTACGTGGTCGAGCTGGACTGGTACCCTGATGCCCCAGGCGAGATGGTC GTGCTGACCTGCGATACACCAGAGGAAGATGGTATCACCTGGACACTGGATCAGT CCTCAGAGGTGCTGGGCTCTGGTAAAACACTGACCATTCAGGTGAAGGAGTTCGG TGACGCTGGACAGTACACTTGTCATAAGGGCGGGGAGGTGCTGTCTCACTCCCTG CTGCTGCTGCATAAGAAGGAGGATGGAATCTGGTCCACTGACATCCTGAAAGACC AGAAGGAGCCAAAGAACAAAACCTTCCTGCGATGCGAGGCTAAGAACTACAGCG GCCGCTTTACATGCTGGTGGCTGACAACCATCAGCACCGATCTGACCTTTAGCGTG AAGTCATCCAGGGGCAGTTCAGACCCTCAGGGAGTCACATGTGGCGCCGCAACC CTGTCAGCAGAGCGAGTGCGGGGAGACAATAAGGAATACGAGTACAGCGTCGAG TGTCAGGAGGATTCCGCATGTCCAGCTGCAGAAGAATCCCTGCCTATCGAAGTCAT GGTGGACGCTGTGCATAAACTGAAGTACGAGAATTACACCAGCAGCTTTTTCATCC GGGACATCATCAAGCCCGATCCACCTAAGAATCTGCAGCTGAAGCCTCTGAAAAA TAGCCGACAGGTCGAAGTGTCATGGGAATACCCAGACACCTGGTCAACACCACAC TCCTACTTCTCCCTGACCTTCTGTGTGCAGGTCCAGGGAAAAAGCAAGCGGGAAA AGAAAGATCGGGTGTTCACCGACAAGACCAGTGCTACAGTGATTTGCCGGAAGAA TGCCAGCATTTCTGTCAGAGCTCAGGACCGGTACTATAGCTCTTCCTGGAGCGAGT GGGCTTCAGTGCCATGTTCTGGAGGCGGTGGATCTGGCGGAGGTGGAAGCGGAG GCGGTGGATCTAGAAACCTGCCCGTCGCAACCCCTGATCCAGGGATGTTCCCCTG TCTGCATCACAGCCAGAATCTGCTGAGGGCTGTCTCCAACATGCTGCAGAAGGCT CGACAGACCCTGGAGTTCTACCCATGTACCAGCGAAGAGATCGACCACGAGGATA TCACAAAGGATAAAACCAGCACAGTGGAAGCATGCCTGCCTCTGGAACTGACCA AGAATGAGAGCTGCCTGAATAGCAGGGAGACCTCCTTCATCACCAACGGCTCATG CCTGGCTTCAAGGAAGACCAGCTTCATGATGGCTCTGTGTCTGAGCTCTATCTATG AGGACCTGAAGATGTACCAGGTGGAGTTCAAGACCATGAACGCCAAGCTGCTGAT GGATCCAAAGAGGCAGATCTTCCTGGATCAGAATATGCTGGCAGTGATCGATGAG CTGATGCAGGCCCTGAATTTTAACAGTGAGACAGTGCCTCAGAAGAGCTCTCTGG AAGAGCCAGACTTTTACAAAACTAAGATCAAGCTGTGCATTCTGCTGCACGCTTT CCGCATCAGAGCTGTCACTATCGATAGAGTGATGAGCTATCTGAATGCCTCA Amino acid sequence of polypeptide chain H8a (SEQ ID NO: 44) EVQLQQSGPGLVKPSQTLSLTCAISGDSVSSNSAAWNWIRQSPSRGLEWLGKTYY RFKWYSDYAVSVKGRITINPDTSKNQFSLQLNSVTPEDTAVFYCTRESTTYDLLAGPF DYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKS CDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNW YVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIE KTISKAKGQPREPQVYTLPPCREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGS GGGGSGGGGSGGGGSIWELKKDVYVVELDWYPDAPGEMVVLTCDTPEEDGITWTL DQSSEVLGSGKTLTIQVKEFGDAGQYTCHKGGEVLSHSLLLLHKKEDGIWSTDILKD QKEPKNKTFLRCEAKNYSGRFTCWWLTTISTDLTFSVKSSRGSSDPQGVTCGAATLSA ERVRGDNKEYEYSVECQEDSACPAAEESLPIEVMVDAVHKLKYENYTSSFFIRDIIKP DPPKNLQLKPLKNSRQVEVSWEYPDTWSTPHSYFSLTFCVQVQGKSKREKKDRVFT DKTSATVICRKNASISVRAQDRYYSSSWSEWASVPCSGGGGSGGGGSGGGGSRNLPV ATPDPGMFPCLHHSQNLLRAVSNMLQKARQTLEFYPCTSEEIDHEDITKDKTSTVEAC LPLELTKNESCLNSRETSFITNGSCLASRKTSFMMALCLSSIYEDLKMYQVEFKTMNA KLLMDPKRQIFLDQNMLAVIDELMQALNFNSETVPQKSSLEEPDFYKTKIKLCILLHA FRIRAVTIDRVMSYLNAS DNA sequence of polypeptide chain H9a (SEQ ID NO: 45) GAAGTGCAGCTGCAGCAGTCCGGCCCCGGACTGGTCAAACCCTCCCAGACCC TGTCTCTTACCTGCGCCATCTCCGGCGACTCTGTGTCCAGCAACTCTGCCGCTTGG AATTGGATCCGGCAGTCTCCTAGTAGAGGCCTGGAATGGCTGGGCAAGACATACT ACCGGTTCAAGTGGTATTCCGATTACGCCGTGTCCGTGAAGGGCAGAATCACCATC AACCCTGATACCTCTAAGAATCAATTCTCCCTGCAGCTGAACTCCGTGACCCCTGA GGACACCGCTGTGTTTTACTGCACCAGAGAGAGCACCACCTACGACCTGCTGGCT GGCCCTTTCGACTACTGGGGCCAGGGAACACTCGTCACCGTGTCTTCCGCCTCTA CAAAGGGCCCATCCGTTTTTCCTCTGGCCCCGTCCTCTAAGTCTACCTCTGGCGGC ACCGCCGCTCTGGGCTGCCTGGTGAAGGACTACTTCCCCGAGCCTGTGACCGTGT CCTGGAACAGCGGCGCTCTGACCTCCGGCGTGCATACCTTCCCTGCTGTTCTGCAA TCTTCTGGCCTGTACTCTCTGTCCTCTGTGGTGACCGTGCCTTCCAGCTCTCTGGG CACCCAGACCTACATCTGCAACGTGAACCATAAGCCTTCTAACACTAAGGTGGAC AAGAAAGTGGAACCCAAGTCCTGCGACAAAACCCACACCTGTCCTCCTTGTCCTG CCCCTGAGCTGCTGGGCGGACCTAGCGTGTTCCTGTTCCCACCTAAGCCTAAGGAT ACCCTGATGATCTCCCGGACCCCTGAAGTGACCTGCGTGGTCGTGGACGTGTCTC ACGAAGATCCTGAAGTCAAGTTCAACTGGTACGTGGATGGCGTCGAGGTGCACAA CGCCAAGACCAAGCCCCGGGAAGAGCAGTACAACTCCACATATAGAGTGGTGAG TGTGCTGACAGTACTGCACCAGGACTGGCTGAACGGCAAAGAGTACAAGTGCAA GGTGTCCAACAAGGCTCTGCCTGCTCCTATCGAGAAGACAATCTCCAAGGCCAAG GGACAGCCTAGAGAACCTCAAGTGTGCACCCTCCCACCTTCTCGCGAGGAGATGA CCAAGAACCAGGTGAGCCTGTCCTGTGCTGTGAAAGGCTTCTACCCCTCCGACAT CGCCGTGGAGTGGGAGTCCAATGGCCAGCCAGAGAACAACTACAAGACCACACC TCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTGGTGTCCAAGCTGACCGTGGAC AAGTCTAGATGGCAGCAGGGCAACGTGTTTAGCTGCTCCGTGATGCACGAGGCCC TGCACAACCACTACACCCAGAAGTCTCTGTCCCTGAGCCCTGGATCAGGTGGAGG CGGTAGTGGCGGAGGCGGTTCAGGCGGAGGCGGATCTGCACCTACTTCAAGTTCT ACAAAGAAAACACAGCTACAACTGGAGCATTTACTGCTGGATTTACAGATGATTTT GAATGGAATTAATAATTACAAGAATCCCAAACTCACCAGGATGCTCACATTTAAGT TTTACATGCCCAAGAAGGCCACAGAACTGAAACATCTTCAGTGTCTAGAAGAAGA ACTCAAACCTCTGGAGGAAGTGCTAAATTTAGCTCAAAGCAAAAACTTTCACTTA AGACCCAGGGACTTAATCAGCAATATCAACGTAATAGTTCTGGAACTAAAGGGATC TGAAACAACATTCATGTGTGAATATGCTGATGAGACAGCAACCATTGTAGAATTTC TGAACAGATGGATTACCTTTTGTCAAAGCATCATCTCAACACTGACT Amino acid sequence of polypeptide chain H9a (SEQ ID NO: 46) EVQLQQSGPGLVKPSQTLSLTCAISGDSVSSNSAAWNWIRQSPSRGLEWLGKTYY RFKWYSDYAVSVKGRITINPDTSKNQFSLQLNSVTPEDTAVFYCTRESTTYDLLAGPF DYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKS CDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNW YVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIE KTISKAKGQPREPQVCTLPPSREEMTKNQVSLSCAVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGS GGGGSGGGGSGGGGSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTF KFYMPKKATELKHLQCLEEELKPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSE TTFMCEYADETATIVEFLNRWITFCQSIISTLT DNA sequence of polypeptide chain H8b (SEQ ID NO: 47) GAAGTGCAGCTGCAGCAGTCCGGCCCCGGACTGGTCAAACCCTCCCAGACCC TGTCTCTTACCTGCGCCATCTCCGGCGACTCTGTGTCCAGCAACTCTGCCGCTTGG AATTGGATCCGGCAGTCTCCTAGTAGAGGCCTGGAATGGCTGGGCAAGACATACT ACCGGTTCAAGTGGTATTCCGATTACGCCGTGTCCGTGAAGGGCAGAATCACCATC AACCCTGATACCTCTAAGAATCAATTCTCCCTGCAGCTGAACTCCGTGACCCCTGA GGACACCGCTGTGTTTTACTGCACCAGAGAGAGCACCACCTACGACCTGCTGGCT GGCCCTTTCGACTACTGGGGCCAGGGAACACTCGTCACCGTGTCTTCCGCCTCTA CAAAGGGCCCATCCGTTTTTCCTCTGGCCCCGTCCTCTAAGTCTACCTCTGGCGGC ACCGCCGCTCTGGGCTGCCTGGTGAAGGACTACTTCCCCGAGCCTGTGACCGTGT CCTGGAACAGCGGCGCTCTGACCTCCGGCGTGCATACCTTCCCTGCTGTTCTGCAA TCTTCTGGCCTGTACTCTCTGTCCTCTGTGGTGACCGTGCCTTCCAGCTCTCTGGG CACCCAGACCTACATCTGCAACGTGAACCATAAGCCTTCTAACACTAAGGTGGAC AAGAAAGTGGAACCCAAGTCCTGCGACAAAACCCACACCTGTCCTCCTTGTCCTG CCCCTGAGCTGCTGGGCGGACCTAGCGTGTTCCTGTTCCCACCTAAGCCTAAGGAT ACCCTGATGATCTCCCGGACCCCTGAAGTGACCTGCGTGGTCGTGGACGTGTCTC ACGAAGATCCTGAAGTCAAGTTCAACTGGTACGTGGATGGCGTCGAGGTGCACAA CGCCAAGACCAAGCCCCGGGAAGAGCAGTACAACTCCACATATAGAGTGGTGAG TGTGCTGACAGTACTGCACCAGGACTGGCTGAACGGCAAAGAGTACAAGTGCAA GGTGTCCAACAAGGCTCTGCCTGCTCCTATCGAGAAGACAATCTCCAAGGCCAAG GGACAGCCTAGAGAACCTCAAGTGTACACCCTCCCACCTTGTCGCGAGGAGATGA CCAAGAACCAGGTGAGCCTGTGGTGTCTGGTGAAAGGCTTCTACCCCTCCGACAT CGCCGTGGAGTGGGAGTCCAATGGCCAGCCAGAGAACAACTACAAGACCACACC TCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTGTACTCCAAGCTGACCGTGGAC AAGTCTAGATGGCAGCAGGGCAACGTGTTTAGCTGCTCCGTGATGCACGAGGCCC TGCACAACCACTACACCCAGAAGTCTCTGTCCCTGAGCCCTGGATCAGGTGGAGG CGGTAGTGGCGGAGGCGGTTCAGGCGGAGGCGGATCTGCACCTACTTCAAGTTCT ACAAAGAAAACACAGCTACAACTGGAGCATTTACTGCTGGATTTACAGATGATTTT GAATGGAATTAATAATTACAAGAATCCCAAACTCACCAGGATGCTCACATTTAAGT TTTACATGCCCAAGAAGGCCACAGAACTGAAACATCTTCAGTGTCTAGAAGAAGA ACTCAAACCTCTGGAGGAAGTGCTAAATTTAGCTCAAAGCAAAAACTTTCACTTA AGACCCAGGGACTTAATCAGCAATATCAACGTAATAGTTCTGGAACTAAAGGGATC TGAAACAACATTCATGTGTGAATATGCTGATGAGACAGCAACCATTGTAGAATTTC TGAACAGATGGATTACCTTTTGTCAAAGCATCATCTCAACACTGACT Amino acid sequence of polypeptide chain H8b (SEQ ID NO: 48) EVQLQQSGPGLVKPSQTLSLTCAISGDSVSSNSAAWNWIRQSPSRGLEWLGKTYY RFKWYSDYAVSVKGRITINPDTSKNQFSLQLNSVTPEDTAVFYCTRESTTYDLLAGPF DYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKS CDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNW YVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIE KTISKAKGQPREPQVYTLPPCREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGS GGGGSGGGGSGGGGSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTF KFYMPKKATELKHLQCLEEELKPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSE TTFMCEYADETATIVEFLNRWITFCQSIISTLT DNA sequence of polypeptide chain H10 (SEQ ID NO: 49) GAAGTGCAGCTGGTGGAATCTGGCGGAGGACTGGTTCAACCTGGCGGCTCTC TGAGACTGTCTTGTGCCGCCAGCGGCTTCGTGTTCAGCAGATACGATATGGCCTGG GTCCGACAGGCCCCTGGCAAAGGACTTGAGTGGGTGTCCTTTATCAGCGGCGGAG GCAGCAACACCTACTATCCCGATACCGTGAAGGGCAGATTCACCATCAGCCGGGA CAACAGCAAGAACACCCTGTACCTGCAGATGAACAGCCTGAGAGCCGAGGACAC CGCCGTGTACTACTGCATCAGCCCCTACTACTACGCCATGGAATACTGGGGCCAGG GCACCACCGTGACAGTGTCTAGCGCCTCTACAAAGGGCCCCAGCGTTTTCCCACT GGCTCCCTGTAGCAGAAGCACCAGCGAATCTACAGCCGCTCTGGGCTGCCTGGTC AAGGACTACTTTCCTGAGCCTGTGACCGTGTCCTGGAACTCTGGCGCTCTGACAA GCGGAGTGCACACCTTTCCAGCCGTGCTGCAAAGCAGCGGCCTGTACTCTCTGAG CAGCGTGGTCACAGTGCCTAGCTCTAGCCTGGGCACCAAGACCTACACCTGTAAT GTGGACCACAAGCCTAGCAACACCAAGGTGGACAAGCGCGTGGAATCTAAGTAC GGCCCTCCTTGTCCTCCATGTCCTGCACCTGAGTTTCTCGGCGGACCCTCCGTGTT CCTGTTTCCTCCAAAGCCTAAGGATACCCTGATGATCAGCAGAACCCCTGAAGTG ACCTGCGTGGTGGTGGACGTGTCCCAAGAGGATCCTGAGGTGCAGTTCAATTGGT ACGTGGACGGCGTGGAAGTGCACAACGCCAAGACCAAGCCTAGAGAGGAACAG TTCAACAGCACCTACAGAGTGGTGTCCGTGCTGACAGTGCTGCACCAGGATTGGC TGAACGGCAAAGAGTACAAGTGCAAGGTGTCCAACAAGGGCCTGCCTAGCAGCA TCGAGAAAACCATCAGCAAGGCCAAGGGCCAGCCAAGAGAACCCCAGGTGTACA CACTGCCTCCAAGCCAAGAGGAAATGACCAAGAACCAGGTGTCCCTGACCTGCC TCGTGAAGGGATTCTACCCCTCCGATATCGCCGTGGAATGGGAGAGCAATGGACA GCCCGAGAACAACTACAAGACAACCCCTCCTGTGCTGGACAGCGACGGCTCATTC TTCCTGTACAGCAGACTGACCGTGGACAAGAGCAGATGGCAAGAGGGCAACGTG TTCTCCTGCAGCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGTCTC TGTCCCTGTCTCTGGGCAAG Amino acid sequence of polypeptide chain H10 (SEQ ID NO: 50) EVQLVESGGGLVQPGGSLRLSCAASGFVFSRYDMAWVRQAPGKGLEWVSFISGG GSNTYYPDTVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCISPYYYAMEYWGQG TTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVH TFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCP APEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNA KTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPR EPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSD GSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK DNA sequence of polypeptide chain L4 (SEQ ID NO: 51) GACATCCAGATGACACAGAGCCCTAGCAGCGTGTCCGCCTCTGTGGGAGACA GAGTGACCATCACATGCAAGGCCAGCCAGGATGTGGATACAGCCGTGGCCTGGTA TCAGCAGAAGCCTGGAAAGGCCCCTAAGCTGCTGATCTACTGGGCCAGCACAAG ACACACAGGCGTGCCCAGCAGATTTTCTGGCAGCGGCTCTGGCACCGACTTCACC CTGACCATATCTAGCCTGCAGCCTGAGGACTTCGCCACCTACTACTGCCAGCAGTA CAGCACATTCCCCTGGACCTTTGGCGGAGGCACCAAGGTGGAAATCAAGCGGAC AGTGGCCGCTCCTAGCGTGTTCATCTTTCCACCTAGCGACGAGCAGCTGAAGTCC GGCACAGCCTCTGTTGTGTGCCTGCTGAACAACTTCTACCCCAGAGAAGCCAAGG TGCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGCAATAGCCAAGAGTCTGTGA CCGAGCAGGACAGCAAGGACTCTACCTACAGCCTGTCCAGCACACTGACCCTGA GCAAGGCCGACTACGAGAAGCACAAAGTGTACGCCTGCGAAGTGACCCACCAGG GCCTTTCTAGCCCTGTGACCAAGAGCTTCAACCGGGGCGAATGT Amino acid sequence of polypeptide chain L4 (SEQ ID NO: 52) DIQMTQSPSSVSASVGDRVTITCKASQDVDTAVAWYQQKPGKAPKLLIYWASTR HTGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYSTFPWTFGGGTKVEIKRTVAAP SVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKD STYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC DNA sequence of polypeptide chain H11 (SEQ ID NO: 53) GAAGTGCAGCTGGTGGAATCTGGCGGAGGACTGGTTCAACCTGGCGGCTCTC TGAGACTGTCTTGTGCCGCCAGCGGCTTCGTGTTCAGCAGATACGATATGGCCTGG GTCCGACAGGCCCCTGGCAAAGGACTTGAGTGGGTGTCCTTTATCAGCGGCGGAG GCAGCAACACCTACTATCCCGATACCGTGAAGGGCAGATTCACCATCAGCCGGGA CAACAGCAAGAACACCCTGTACCTGCAGATGAACAGCCTGAGAGCCGAGGACAC CGCCGTGTACTACTGCATCAGCCCCTACTACTACGCCATGGAATACTGGGGCCAGG GCACCACCGTGACAGTGTCTAGCGCCTCTACAAAGGGCCCCAGCGTTTTCCCACT GGCTCCCTGTAGCAGAAGCACCAGCGAATCTACAGCCGCTCTGGGCTGCCTGGTC AAGGACTACTTTCCTGAGCCTGTGACCGTGTCCTGGAACTCTGGCGCTCTGACAA GCGGAGTGCACACCTTTCCAGCCGTGCTGCAAAGCAGCGGCCTGTACTCTCTGAG CAGCGTGGTCACAGTGCCTAGCTCTAGCCTGGGCACCAAGACCTACACCTGTAAT GTGGACCACAAGCCTAGCAACACCAAGGTGGACAAGCGCGTGGAATCTAAGTAC GGCCCTCCTTGTCCTCCATGTCCTGCACCTGAGTTTCTCGGCGGACCCTCCGTGTT CCTGTTTCCTCCAAAGCCTAAGGATACCCTGATGATCAGCAGAACCCCTGAAGTG ACCTGCGTGGTGGTGGACGTGTCCCAAGAGGATCCTGAGGTGCAGTTCAATTGGT ACGTGGACGGCGTGGAAGTGCACAACGCCAAGACCAAGCCTAGAGAGGAACAG TTCAACAGCACCTACAGAGTGGTGTCCGTGCTGACAGTGCTGCACCAGGATTGGC TGAACGGCAAAGAGTACAAGTGCAAGGTGTCCAACAAGGGCCTGCCTAGCAGCA TCGAGAAAACCATCAGCAAGGCCAAGGGCCAGCCAAGAGAACCCCAGGTGTACA CACTGCCTCCATGCCAAGAGGAAATGACCAAGAACCAGGTGTCCCTGTGGTGCCT CGTGAAGGGATTCTACCCCTCCGATATCGCCGTGGAATGGGAGAGCAATGGACAG CCCGAGAACAACTACAAGACAACCCCTCCTGTGCTGGACAGCGACGGCTCATTCT TCCTGTACAGCAGACTGACCGTGGACAAGAGCAGATGGCAAGAGGGCAACGTGT TCTCCTGCAGCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGTCTCT GTCCCTGTCTCTGGGC Amino acid sequence of polypeptide chain H11 (SEQ ID NO: 54) EVQLVESGGGLVQPGGSLRLSCAASGFVFSRYDMAWVRQAPGKGLEWVSFISGG GSNTYYPDTVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCISPYYYAMEYWGQG TTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVH TFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCP APEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNA KTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPR EPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSD GSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG DNA sequence of polypeptide chain H12 (SEQ ID NO: 55) GAAGTGCAGCTGGTGGAATCTGGCGGAGGACTGGTTCAACCTGGCGGCTCTC TGAGACTGTCTTGTGCCGCCAGCGGCTTCGTGTTCAGCAGATACGATATGGCCTGG GTCCGACAGGCCCCTGGCAAAGGACTTGAGTGGGTGTCCTTTATCAGCGGCGGAG GCAGCAACACCTACTATCCCGATACCGTGAAGGGCAGATTCACCATCAGCCGGGA CAACAGCAAGAACACCCTGTACCTGCAGATGAACAGCCTGAGAGCCGAGGACAC CGCCGTGTACTACTGCATCAGCCCCTACTACTACGCCATGGAATACTGGGGCCAGG GCACCACCGTGACAGTGTCTAGCGCCTCTACAAAGGGCCCCAGCGTTTTCCCACT GGCTCCCTGTAGCAGAAGCACCAGCGAATCTACAGCCGCTCTGGGCTGCCTGGTC AAGGACTACTTTCCTGAGCCTGTGACCGTGTCCTGGAACTCTGGCGCTCTGACAA GCGGAGTGCACACCTTTCCAGCCGTGCTGCAAAGCAGCGGCCTGTACTCTCTGAG CAGCGTGGTCACAGTGCCTAGCTCTAGCCTGGGCACCAAGACCTACACCTGTAAT GTGGACCACAAGCCTAGCAACACCAAGGTGGACAAGCGCGTGGAATCTAAGTAC GGCCCTCCTTGTCCTCCATGTCCTGCACCTGAGTTTCTCGGCGGACCCTCCGTGTT CCTGTTTCCTCCAAAGCCTAAGGATACCCTGATGATCAGCAGAACCCCTGAAGTG ACCTGCGTGGTGGTGGACGTGTCCCAAGAGGATCCTGAGGTGCAGTTCAATTGGT ACGTGGACGGCGTGGAAGTGCACAACGCCAAGACCAAGCCTAGAGAGGAACAG TTCAACAGCACCTACAGAGTGGTGTCCGTGCTGACAGTGCTGCACCAGGATTGGC TGAACGGCAAAGAGTACAAGTGCAAGGTGTCCAACAAGGGCCTGCCTAGCAGCA TCGAGAAAACCATCAGCAAGGCCAAGGGCCAGCCAAGAGAACCCCAGGTGTGCA CACTGCCTCCAAGCCAAGAGGAAATGACCAAGAACCAGGTGTCCCTGTCCTGCG CCGTGAAGGGATTCTACCCCTCCGATATCGCCGTGGAATGGGAGAGCAATGGACA GCCCGAGAACAACTACAAGACAACCCCTCCTGTGCTGGACAGCGACGGCTCATTC TTCCTGGTGAGCAGACTGACCGTGGACAAGAGCAGATGGCAAGAGGGCAACGTG TTCTCCTGCAGCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGTCTC TGTCCCTGTCTCTGGGC Amino acid sequence of polypeptide chain H12 (SEQ ID NO: 56) EVQLVESGGGLVQPGGSLRLSCAASGFVFSRYDMAWVRQAPGKGLEWVSFISGG GSNTYYPDTVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCISPYYYAMEYWGQG TTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVH TFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCP APEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNA KTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPR EPQVCTLPPSQEEMTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSD GSFFLVSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG DNA sequence of polypeptide chain H11a (SEQ ID NO: 57) GAAGTGCAGCTGGTGGAATCTGGCGGAGGACTGGTTCAACCTGGCGGCTCTC TGAGACTGTCTTGTGCCGCCAGCGGCTTCGTGTTCAGCAGATACGATATGGCCTGG GTCCGACAGGCCCCTGGCAAAGGACTTGAGTGGGTGTCCTTTATCAGCGGCGGAG GCAGCAACACCTACTATCCCGATACCGTGAAGGGCAGATTCACCATCAGCCGGGA CAACAGCAAGAACACCCTGTACCTGCAGATGAACAGCCTGAGAGCCGAGGACAC CGCCGTGTACTACTGCATCAGCCCCTACTACTACGCCATGGAATACTGGGGCCAGG GCACCACCGTGACAGTGTCTAGCGCCTCTACAAAGGGCCCCAGCGTTTTCCCACT GGCTCCCTGTAGCAGAAGCACCAGCGAATCTACAGCCGCTCTGGGCTGCCTGGTC AAGGACTACTTTCCTGAGCCTGTGACCGTGTCCTGGAACTCTGGCGCTCTGACAA GCGGAGTGCACACCTTTCCAGCCGTGCTGCAAAGCAGCGGCCTGTACTCTCTGAG CAGCGTGGTCACAGTGCCTAGCTCTAGCCTGGGCACCAAGACCTACACCTGTAAT GTGGACCACAAGCCTAGCAACACCAAGGTGGACAAGCGCGTGGAATCTAAGTAC GGCCCTCCTTGTCCTCCATGTCCTGCACCTGAGTTTCTCGGCGGACCCTCCGTGTT CCTGTTTCCTCCAAAGCCTAAGGATACCCTGATGATCAGCAGAACCCCTGAAGTG ACCTGCGTGGTGGTGGACGTGTCCCAAGAGGATCCTGAGGTGCAGTTCAATTGGT ACGTGGACGGCGTGGAAGTGCACAACGCCAAGACCAAGCCTAGAGAGGAACAG TTCAACAGCACCTACAGAGTGGTGTCCGTGCTGACAGTGCTGCACCAGGATTGGC TGAACGGCAAAGAGTACAAGTGCAAGGTGTCCAACAAGGGCCTGCCTAGCAGCA TCGAGAAAACCATCAGCAAGGCCAAGGGCCAGCCAAGAGAACCCCAGGTGTACA CACTGCCTCCATGCCAAGAGGAAATGACCAAGAACCAGGTGTCCCTGTGGTGCCT CGTGAAGGGATTCTACCCCTCCGATATCGCCGTGGAATGGGAGAGCAATGGACAG CCCGAGAACAACTACAAGACAACCCCTCCTGTGCTGGACAGCGACGGCTCATTCT TCCTGTACAGCAGACTGACCGTGGACAAGAGCAGATGGCAAGAGGGCAACGTGT TCTCCTGCAGCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGTCTCT GTCCCTGTCTCTGGGCTCAGGTGGAGGCGGTAGTGGCGGAGGCGGTTCAGGCGG AGGCGGATCTATTTGGGAGCTGAAGAAAGACGTGTACGTGGTCGAGCTGGACTGG TACCCTGATGCCCCAGGCGAGATGGTCGTGCTGACCTGCGATACACCAGAGGAAG ATGGTATCACCTGGACACTGGATCAGTCCTCAGAGGTGCTGGGCTCTGGTAAAAC ACTGACCATTCAGGTGAAGGAGTTCGGTGACGCTGGACAGTACACTTGTCATAAG GGCGGGGAGGTGCTGTCTCACTCCCTGCTGCTGCTGCATAAGAAGGAGGATGGAA TCTGGTCCACTGACATCCTGAAAGACCAGAAGGAGCCAAAGAACAAAACCTTCC TGCGATGCGAGGCTAAGAACTACAGCGGCCGCTTTACATGCTGGTGGCTGACAAC CATCAGCACCGATCTGACCTTTAGCGTGAAGTCATCCAGGGGCAGTTCAGACCCT CAGGGAGTCACATGTGGCGCCGCAACCCTGTCAGCAGAGCGAGTGCGGGGAGAC AATAAGGAATACGAGTACAGCGTCGAGTGTCAGGAGGATTCCGCATGTCCAGCTG CAGAAGAATCCCTGCCTATCGAAGTCATGGTGGACGCTGTGCATAAACTGAAGTA CGAGAATTACACCAGCAGCTTTTTCATCCGGGACATCATCAAGCCCGATCCACCTA AGAATCTGCAGCTGAAGCCTCTGAAAAATAGCCGACAGGTCGAAGTGTCATGGGA ATACCCAGACACCTGGTCAACACCACACTCCTACTTCTCCCTGACCTTCTGTGTGC AGGTCCAGGGAAAAAGCAAGCGGGAAAAGAAAGATCGGGTGTTCACCGACAAG ACCAGTGCTACAGTGATTTGCCGGAAGAATGCCAGCATTTCTGTCAGAGCTCAGG ACCGGTACTATAGCTCTTCCTGGAGCGAGTGGGCTTCAGTGCCATGTTCTGGAGGC GGTGGATCTGGCGGAGGTGGAAGCGGAGGCGGTGGATCTAGAAACCTGCCCGTC GCAACCCCTGATCCAGGGATGTTCCCCTGTCTGCATCACAGCCAGAATCTGCTGA GGGCTGTCTCCAACATGCTGCAGAAGGCTCGACAGACCCTGGAGTTCTACCCATG TACCAGCGAAGAGATCGACCACGAGGATATCACAAAGGATAAAACCAGCACAGT GGAAGCATGCCTGCCTCTGGAACTGACCAAGAATGAGAGCTGCCTGAATAGCAGG GAGACCTCCTTCATCACCAACGGCTCATGCCTGGCTTCAAGGAAGACCAGCTTCA TGATGGCTCTGTGTCTGAGCTCTATCTATGAGGACCTGAAGATGTACCAGGTGGAG TTCAAGACCATGAACGCCAAGCTGCTGATGGATCCAAAGAGGCAGATCTTCCTGG ATCAGAATATGCTGGCAGTGATCGATGAGCTGATGCAGGCCCTGAATTTTAACAGT GAGACAGTGCCTCAGAAGAGCTCTCTGGAAGAGCCAGACTTTTACAAAACTAAG ATCAAGCTGTGCATTCTGCTGCACGCTTTCCGCATCAGAGCTGTCACTATCGATAG AGTGATGAGCTATCTGAATGCCTCA Amino acid sequence of polypeptide chain H11a (SEQ ID NO: 58) EVQLVESGGGLVQPGGSLRLSCAASGFVFSRYDMAWVRQAPGKGLEWVSFISGG GSNTYYPDTVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCISPYYYAMEYWGQG TTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVH TFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCP APEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNA KTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPR EPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSD GSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGSGGGGSGGGGS GGGGSIWELKKDVYVVELDWYPDAPGEMVVLTCDTPEEDGITWTLDQSSEVLGSGK TLTIQVKEFGDAGQYTCHKGGEVLSHSLLLLHKKEDGIWSTDILKDQKEPKNKTFLR CEAKNYSGRFTCWWLTTISTDLTFSVKSSRGSSDPQGVTCGAATLSAERVRGDNKEY EYSVECQEDSACPAAEESLPIEVMVDAVHKLKYENYTSSFFIRDIIKPDPPKNLQLKPL KNSRQVEVSWEYPDTWSTPHSYFSLTFCVQVQGKSKREKKDRVFTDKTSATVICRKN ASISVRAQDRYYSSSWSEWASVPCSGGGGSGGGGSGGGGSRNLPVATPDPGMFPCLH HSQNLLRAVSNMLQKARQTLEFYPCTSEEIDHEDITKDKTSTVEACLPLELTKNESCL NSRETSFITNGSCLASRKTSFMMALCLSSIYEDLKMYQVEFKTMNAKLLMDPKRQIF LDQNMLAVIDELMQALNFNSETVPQKSSLEEPDFYKTKIKLCILLHAFRIRAVTIDRVM SYLNAS DNA sequence of polypeptide chain H12a (SEQ ID NO: 59) GAAGTGCAGCTGGTGGAATCTGGCGGAGGACTGGTTCAACCTGGCGGCTCTC TGAGACTGTCTTGTGCCGCCAGCGGCTTCGTGTTCAGCAGATACGATATGGCCTGG GTCCGACAGGCCCCTGGCAAAGGACTTGAGTGGGTGTCCTTTATCAGCGGCGGAG GCAGCAACACCTACTATCCCGATACCGTGAAGGGCAGATTCACCATCAGCCGGGA CAACAGCAAGAACACCCTGTACCTGCAGATGAACAGCCTGAGAGCCGAGGACAC CGCCGTGTACTACTGCATCAGCCCCTACTACTACGCCATGGAATACTGGGGCCAGG GCACCACCGTGACAGTGTCTAGCGCCTCTACAAAGGGCCCCAGCGTTTTCCCACT GGCTCCCTGTAGCAGAAGCACCAGCGAATCTACAGCCGCTCTGGGCTGCCTGGTC AAGGACTACTTTCCTGAGCCTGTGACCGTGTCCTGGAACTCTGGCGCTCTGACAA GCGGAGTGCACACCTTTCCAGCCGTGCTGCAAAGCAGCGGCCTGTACTCTCTGAG CAGCGTGGTCACAGTGCCTAGCTCTAGCCTGGGCACCAAGACCTACACCTGTAAT GTGGACCACAAGCCTAGCAACACCAAGGTGGACAAGCGCGTGGAATCTAAGTAC GGCCCTCCTTGTCCTCCATGTCCTGCACCTGAGTTTCTCGGCGGACCCTCCGTGTT CCTGTTTCCTCCAAAGCCTAAGGATACCCTGATGATCAGCAGAACCCCTGAAGTG ACCTGCGTGGTGGTGGACGTGTCCCAAGAGGATCCTGAGGTGCAGTTCAATTGGT ACGTGGACGGCGTGGAAGTGCACAACGCCAAGACCAAGCCTAGAGAGGAACAG TTCAACAGCACCTACAGAGTGGTGTCCGTGCTGACAGTGCTGCACCAGGATTGGC TGAACGGCAAAGAGTACAAGTGCAAGGTGTCCAACAAGGGCCTGCCTAGCAGCA TCGAGAAAACCATCAGCAAGGCCAAGGGCCAGCCAAGAGAACCCCAGGTGTGCA CACTGCCTCCAAGCCAAGAGGAAATGACCAAGAACCAGGTGTCCCTGTCCTGCG CCGTGAAGGGATTCTACCCCTCCGATATCGCCGTGGAATGGGAGAGCAATGGACA GCCCGAGAACAACTACAAGACAACCCCTCCTGTGCTGGACAGCGACGGCTCATTC TTCCTGGTGAGCAGACTGACCGTGGACAAGAGCAGATGGCAAGAGGGCAACGTG TTCTCCTGCAGCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGTCTC TGTCCCTGTCTCTGGGCTCAGGTGGAGGCGGTAGTGGCGGAGGCGGTTCAGGCG GAGGCGGATCTGCACCTACTTCAAGTTCTACAAAGAAAACACAGCTACAACTGGA GCATTTACTGCTGGATTTACAGATGATTTTGAATGGAATTAATAATTACAAGAATCC CAAACTCACCAGGATGCTCACATTTAAGTTTTACATGCCCAAGAAGGCCACAGAA CTGAAACATCTTCAGTGTCTAGAAGAAGAACTCAAACCTCTGGAGGAAGTGCTAA ATTTAGCTCAAAGCAAAAACTTTCACTTAAGACCCAGGGACTTAATCAGCAATATC AACGTAATAGTTCTGGAACTAAAGGGATCTGAAACAACATTCATGTGTGAATATGC TGATGAGACAGCAACCATTGTAGAATTTCTGAACAGATGGATTACCTTTTGTCAAA GCATCATCTCAACACTGACT Amino acid sequence of polypeptide chain H12a (SEQ ID NO: 60) EVQLVESGGGLVQPGGSLRLSCAASGFVFSRYDMAWVRQAPGKGLEWVSFISGG GSNTYYPDTVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCISPYYYAMEYWGQG TTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVH TFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCP APEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNA KTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPR EPQVCTLPPSQEEMTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSD GSFFLVSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGSGGGGSGGGGS GGGGSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTFKFYMPKKATE LKHLQCLEEELKPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADET ATIVEFLNRWITFCQSIISTLT DNA sequence of polypeptide chain H11b (SEQ ID NO: 61) GAAGTGCAGCTGGTGGAATCTGGCGGAGGACTGGTTCAACCTGGCGGCTCTC TGAGACTGTCTTGTGCCGCCAGCGGCTTCGTGTTCAGCAGATACGATATGGCCTGG GTCCGACAGGCCCCTGGCAAAGGACTTGAGTGGGTGTCCTTTATCAGCGGCGGAG GCAGCAACACCTACTATCCCGATACCGTGAAGGGCAGATTCACCATCAGCCGGGA CAACAGCAAGAACACCCTGTACCTGCAGATGAACAGCCTGAGAGCCGAGGACAC CGCCGTGTACTACTGCATCAGCCCCTACTACTACGCCATGGAATACTGGGGCCAGG GCACCACCGTGACAGTGTCTAGCGCCTCTACAAAGGGCCCCAGCGTTTTCCCACT GGCTCCCTGTAGCAGAAGCACCAGCGAATCTACAGCCGCTCTGGGCTGCCTGGTC AAGGACTACTTTCCTGAGCCTGTGACCGTGTCCTGGAACTCTGGCGCTCTGACAA GCGGAGTGCACACCTTTCCAGCCGTGCTGCAAAGCAGCGGCCTGTACTCTCTGAG CAGCGTGGTCACAGTGCCTAGCTCTAGCCTGGGCACCAAGACCTACACCTGTAAT GTGGACCACAAGCCTAGCAACACCAAGGTGGACAAGCGCGTGGAATCTAAGTAC GGCCCTCCTTGTCCTCCATGTCCTGCACCTGAGTTTCTCGGCGGACCCTCCGTGTT CCTGTTTCCTCCAAAGCCTAAGGATACCCTGATGATCAGCAGAACCCCTGAAGTG ACCTGCGTGGTGGTGGACGTGTCCCAAGAGGATCCTGAGGTGCAGTTCAATTGGT ACGTGGACGGCGTGGAAGTGCACAACGCCAAGACCAAGCCTAGAGAGGAACAG TTCAACAGCACCTACAGAGTGGTGTCCGTGCTGACAGTGCTGCACCAGGATTGGC TGAACGGCAAAGAGTACAAGTGCAAGGTGTCCAACAAGGGCCTGCCTAGCAGCA TCGAGAAAACCATCAGCAAGGCCAAGGGCCAGCCAAGAGAACCCCAGGTGTACA CACTGCCTCCATGCCAAGAGGAAATGACCAAGAACCAGGTGTCCCTGTGGTGCCT CGTGAAGGGATTCTACCCCTCCGATATCGCCGTGGAATGGGAGAGCAATGGACAG CCCGAGAACAACTACAAGACAACCCCTCCTGTGCTGGACAGCGACGGCTCATTCT TCCTGTACAGCAGACTGACCGTGGACAAGAGCAGATGGCAAGAGGGCAACGTGT TCTCCTGCAGCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGTCTCT GTCCCTGTCTCTGGGCTCAGGTGGAGGCGGTAGTGGCGGAGGCGGTTCAGGCGG AGGCGGATCTGCACCTACTTCAAGTTCTACAAAGAAAACACAGCTACAACTGGAG CATTTACTGCTGGATTTACAGATGATTTTGAATGGAATTAATAATTACAAGAATCCC AAACTCACCAGGATGCTCACATTTAAGTTTTACATGCCCAAGAAGGCCACAGAAC TGAAACATCTTCAGTGTCTAGAAGAAGAACTCAAACCTCTGGAGGAAGTGCTAAA TTTAGCTCAAAGCAAAAACTTTCACTTAAGACCCAGGGACTTAATCAGCAATATCA ACGTAATAGTTCTGGAACTAAAGGGATCTGAAACAACATTCATGTGTGAATATGCT GATGAGACAGCAACCATTGTAGAATTTCTGAACAGATGGATTACCTTTTGTCAAAG CATCATCTCAACACTGACT Amino acid sequence of polypeptide chain H11b (SEQ ID NO: 62) EVQLVESGGGLVQPGGSLRLSCAASGFVFSRYDMAWVRQAPGKGLEWVSFISGG GSNTYYPDTVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCISPYYYAMEYWGQG TTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVH TFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCP APEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNA KTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPR EPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSD GSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGSGGGGSGGGGS GGGGSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTFKFYMPKKATE LKHLQCLEEELKPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADET ATIVEFLNRWITFCQSIISTLT Linker sequence (SEQ ID NO: 63): SGGGGSGGGGSGGGGS 

1. A monoclonal antibody-cytokine fusion protein dimer, wherein a monoclonal antibody comprises two heavy chains, the fusion protein dimer comprises a first heavy chain polypeptide chain and a second heavy chain polypeptide chain, wherein one heavy chain of the monoclonal antibody is connected with a cytokine to form the first heavy chain polypeptide chain, and the other heavy chain of the monoclonal antibody is optionally connected with another cytokine to form the second heavy chain polypeptide chain.
 2. The fusion protein dimer according to claim 1, wherein the monoclonal antibody is an anti-immune checkpoint molecular antibody.
 3. The fusion protein dimer according to claim 1, wherein the monoclonal antibody is selected from an anti-LAG-3 antibody, an anti-PD-L1 antibody, an anti-TIGIT antibody or an anti-PD-1 antibody.
 4. The fusion protein dimer according to claim 1, wherein the two cytokines are selected from IL-2, IL-12, GM-CSF, an IL-2 mutant or a combination thereof.
 5. (canceled)
 6. The fusion protein dimer according to claim 1, wherein the monoclonal antibody is connected with the cytokines through Fc regions of the heavy chains.
 7. The fusion protein dimer according to claim 6, wherein the Fc regions of the two heavy chains of the monoclonal antibody are different and have an asymmetric complementary structure there between.
 8. The fusion protein dimer according to claim 7, wherein the Fc region of any one heavy chain of the monoclonal antibody has a mutation combination T366W/S354C, and the Fc region of the other complementary heavy chain has a mutation combination T366S/L368A/Y407V/Y349C.
 9. The fusion protein dimer according to claim 1, wherein one heavy chain of the monoclonal antibody is connected with the cytokine IL-12 to form the first heavy chain polypeptide chain, and the other heavy chain of the monoclonal antibody is connected with the cytokine IL-2 or the IL-2 mutant to form the second heavy chain polypeptide chain; or one heavy chain of the monoclonal antibody is connected with the cytokine IL-2 or the IL-2 mutant to form the first heavy chain polypeptide chain, and the other heavy chain of the monoclonal antibody is connected with the cytokine IL-12 to form the second heavy chain polypeptide chain.
 10. The fusion protein dimer according to claim 1, wherein one heavy chain of the monoclonal antibody is connected with the cytokine IL-12 or IL-2 to form the first heavy chain polypeptide chain, and the other heavy chain of the monoclonal antibody is not connected with the cytokine to serve as the second heavy chain polypeptide chain.
 11. (canceled)
 12. The fusion protein dimer according to claim 3, wherein a heavy chain of the anti-PD-L1 monoclonal antibody has a sequence with at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identity with an amino acid sequence shown in SEQ ID NOs:22, 26 or 28, and a light chain of the anti-PD-L1 monoclonal antibody has a sequence with at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identity with an amino acid sequence shown in SEQ ID NO:24.
 13. (canceled)
 14. The fusion protein dimer according to claim 3, wherein a heavy chain of the anti-PD-1 monoclonal antibody has a sequence with at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identity with an amino acid sequence shown in SEQ ID NOs:50, 54 or 56, and a light chain of the anti-PD-1 monoclonal antibody has a sequence with at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identity with an amino acid sequence shown in SEQ ID NO:52.
 15. The fusion protein dimer according to claim 4, wherein the IL-12 single strand protein has an amino acid sequence as shown in SEQ ID NO:4. 16.-18. (canceled)
 19. The fusion protein dimer according to claim 1, wherein monoclonal antibody is the anti-PD-1 antibody, and wherein the first heavy chain polypeptide chain has the amino acid sequence as shown in SEQ ID NO:60, and the second heavy chain polypeptide chain has the amino acid sequence as shown in SEQ ID NO:58; the first heavy chain polypeptide chain has the amino acid sequence as shown in SEQ ID NO:58, and the second heavy chain polypeptide chain has the amino acid sequence as shown in SEQ ID NO:56; or the first heavy chain polypeptide chain has the amino acid sequence as shown in SEQ ID NO:62, and the second heavy chain polypeptide chain has the amino acid sequence as shown in SEQ ID NO:56.
 20. The fusion protein dimer according to claim 1, wherein the monoclonal antibody and the cytokines are connected through the linker sequence or directly fused.
 21. The fusion protein dimer according to claim 20, wherein the linker sequence is selected from (S(G4S)₁₋₃), (G4S)₁₋₃, KRVAPELLGGPS, ASTKG, NSPPAA, EPKSSDKTHTSPPSP, ERKSSVESPPSP and ESKYGPPSPPSP.
 22. An isolated polynucleotide, encoding the fusion protein dimer according to claim
 1. 23. An expression vector comprising the polynucleotide according to claim
 22. 24. A host cell comprising the expression vector of claim
 23. 25. A method of treating tumor, comprising administrating to the subject a therapeutically effective amount of the fusion protein dimer according to claim
 1. 26. An anti-tumor pharmaceutical composition, comprising the fusion protein dimer according to claim 1 and a pharmaceutically acceptable carrier. 